http://2006.igem.org/wiki/index.php?title=Special:Contributions/Nadeau&feed=atom&limit=50&target=Nadeau&year=&month=2006.igem.org - User contributions [en]2024-03-28T20:56:46ZFrom 2006.igem.orgMediaWiki 1.16.5http://2006.igem.org/wiki/index.php/May_2007May 20072007-05-14T20:39:36Z<p>Nadeau: </p>
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<div>*May 11<br />
Autoclaved LB, Agar and Minimal Medium. <br />
Made plates with AmpR, KanR and both Amp and Kan. Concentrations used (same for cell cultures):<br />
Amp: 1uL/mL<br />
Kan: 10uL/mL<br />
Cam: 0.8uL/mL (from Elvis' aliquots)<br />
The receipe for M9 Minimal Medium is in my lab notebook which is in the lab so I'll post it later under the Protocols section. Hopefully we can get the 2007 wiki soon. <b>Horia</b><br />
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*May 14<br />
Moved plates from iGEM fridge to fridge on C block (end of hall)<br />
Made 100X Yeast extract and 10X Dextrose (for supplementing minimal medium)<br />
Prepared CaCl2 and CaCl2/Glycerol 10% solutions for CC procedure of tomorrow<br />
Will seed tonight. <b>Horia</b></div>Nadeauhttp://2006.igem.org/wiki/index.php/May_2007May 20072007-05-12T04:32:23Z<p>Nadeau: </p>
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<div>*May 11<br />
Autoclaved LB, Agar and Minimal Medium. <br />
Made plates with AmpR, KanR and both Amp and Kan. Concentrations used (same for cell cultures):<br />
Amp: 1uL/mL<br />
Kan: 10uL/mL<br />
Cam: 0.8uL/mL (from Elvis' aliquots)<br />
The receipe for M9 Minimal Medium is in my lab notebook which is in the lab so I'll post it later under the Protocols section. Hopefully we can get the 2007 wiki soon. <b>Horia</b></div>Nadeauhttp://2006.igem.org/wiki/index.php/Lab_NotebookLab Notebook2007-05-12T04:23:21Z<p>Nadeau: </p>
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<div>[[McGill_University_2006|Home]] <br>[[File_registry|File Registry]]<br />
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[[May|May]] [[June|June]] [[July|July]] [[Team 1|Team 1]] [[May 2007]]<br />
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'''July 1''' Saturday<br />
*Hmmm, I hope people haven't stopped working just because I'm at the north pole... here at the north pole, we have stopped working for a bit, due to bad weather, and are sitting around eating, getting fat, and sending email. Quite the life. There's snow. Hope the ligations are working.<br />
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'''July 3''' Monday<br />
*Team 1: Decided to cut what was isolated from midi-preps w/ BamH1. Going to run digests on a gel to see if we get characteristic bands. Primers should arrive tomorrow for YFP PCR. Hope you're having fun Jay!<br />
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*Team 2: Set up screening digestion of E0422/C0012 ligation made master mix with the following quantities: 3.75 uL enzyme BsrbI, 7.5 uL Buffer2, 48.75 uL dH2O. This was divded into 5 tubes and 3 uL of miniprep DNA was added to each. Ran a gel of the digested DNA and un-digested miniprep DNA and found that there was no DNA. Did DNA precipitation following following protocol on 6/22/06. Prepared screening digestion of the precipitated DNA. Digested overnight.<br />
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*Team 3: Ashwin and Jieun: New plating of the transformed Top 10F containing the ligation product was performed on two amp plates. For the future use, another ligation was performed using the same DNAs (Luc from June 21st, which had very strong bands on the gel analysis and GFP from Elvis) with the same ratio from June 21st.<br />
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'''July 4''' Tuesday<br />
*Team 3: Jieun: Transformation of ligation product was performed (one vial of Top10F with 10uL of ligation product). The other 20uL of ligation product is stored in 4C fridge for overnight, and the plates containing trasformed cells are kept in 37C incubator room (starting from 5:30pm EST).<br />
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*Seeded 2 colonies from pB10+Eco-Mscl in DH5a overnight.<br />
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*Team 2: Ran a gel of our precipitated DNA and digested DNA. Found there was no DNA. Seeded more colonies of the E0422/C0012 ligation.<br />
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*Team 1: Did three seedings with 2 controls in 2 mL. Digested Fos-Beta a second time. Ran Fos-Beta on a gel but had no DNA in our lane. At 5 PM, added more LB to our seedings and left them overnight in incubator. <br />
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'''July 5'''<br />
*Team 2: Miniprepped seedings from yesterday, followed protocol on May 9, 2006. Set up screening digestion of miniprepped DNA. Used the same quantities as on July 3, 2006. Ran a gel of this digested screening and undigested miniprep. All 5 samples had DNA in them. Unable to tell if the DNA was cut properly because the lanes were smeared significanly. Made new TAE IX Buffer in an attempt to fix the constant smearing.<br />
*Team 1: Out of 3 seedings, only one grew. Did 3 mini-preps out of that one seeding. Ran 10 microliters of one miniprep in a gel to test if we got DNA. Nothing showed up. Decided to go back to culture plates. Transferred bacteria from one plate onto another amp plate to test if they are truly amp resistant. Put in 37 degee incubator overnight. Received primers for PCR! Will check plate tomorrow. If colonies grow, we're good. If not, need to consider redoing transformation.<br />
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'''July 6'''<br />
*Team 1: We are good, we got colonies! Yay! Picked 10 off plate and did 10 seedings, each in 2 mL. Also did two digests, one of Fos-Beta DNA and one of miniprep from colonies off original pDsBiFC plate. Also just ran non-digested mini-prep DNA to test hypothesis if we are losing DNA with restriction digests. Ran a gel. The Fos-Beta showed up beautifully with the right size bands so now we know we have isolated both the Fos-Beta and Jun-Beta plasmids! However, no DNA showed up in the lanes for the miniprep. That's ok though because we have 10 new seedings. Came back in the evening to find that all 10 seedings grew and the controls were clear. Spun 10 tubes down and resuspended with P1. In the end, had two tubes of resuspended cells. Saved cells and culture tubes in fridge.<br />
*Team 2: Did large scale digest for each of the 5 miniprep samples from yesterday. Expected bands at 1800 bp and 2030 bp. Ran gel of digests and had a band at 1800 and 500 bp. Came to the conclusion that the ligation product did not work properly. Screening for E0422 original plasmid minipreps: Cut with BsrBI. <br />
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'''July 7'''<br />
*Team 2: Ran gel of E0422 screening digests. Expected bands at 1800 and 1200 bp, well 1 had band at 1800, well 2 had band at 2000, and well 3 had band at 1800, 1600, and 1200 bp. Transformed 2 uL of E0422 brick supplied in homemade Top10 cells.<br />
*Team 1: Completed 2 minipreps from the cultures that grew last night. Then did 2 digests of miniprep DNA w/ EcoRI and BamHI and ran a gel of those digests in the afternoon. Purpose was to screen PLasmid DNA to make sure it is the pDsBiFC plasmid that we want. <br />
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'''July 9'''<br />
*Team 2: Seeded 4 colonies from transformation plate into LB + Amp for miniprep.<br />
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'''July 10 Monday'''<br />
*Team 1: Results from gel that was run on the 7th were not good because it seems the enzymes did not cut. Decided to run three more digests, 2 single ( 1 EcoRI and 1 BamHI, and 1 double w/ both of them) We then ran all digests on the same gel and we got our expected results! Yes! The only problem is the bands were so faint that you could barely see them. <br />
*Team 2: Miniprepped 5 mL of broth culture for each column. Prepared screening digests with BsrBI according to proportions listed on July 6/06. After running on gel, found only sample 4 showed a band, and it was at about 2000 bp. Annette recommended screening with a different enzyme (NotI). Set up screening digest with NotI. Run gel of digests with samples of today's miniprep to see if any DNA present in these minipreps. Well 4 was the only well to contain the correct DNA (band at 3000, 2000, and 1000 bp). Well 6 and 7 contained DNA but the identity could not be confirmed. Well 5 did not contain DNA, and wells 1, 2, and 3, did not contain the correct plasmid.<br />
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'''July 11'''<br />
*Team 2: Complete a large scale digest of the sample run in well 4. Cut using EcoRI, XbaI, and ScaI. Realized that this was very bad, because it destroyed our plasmid. Oops. Re-did the large scale digestion using EcoRI and XbaI, digested for 4 hours. Added 3 uL of CIP, incubated at 37C overnight.<br />
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'''July 12'''<br />
*Team 2: Performed DNA desalting to remove enzymes and small DNA fragment removed between EcoRI and XbaI sites. 53 uL sample volume -> 159 uL QX1, 3.8 ug DNA -> 10 uL QiaexII. Need to ligate C0012 brick (cut at EcoRI and SpeI) into E0422 with plasmid (cut at EcoRI and XbaI). Did calculations for ligation, set up ligation of E0422 and C0012, transformed into commercial Top10, and plated.<br />
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'''July 13'''<br />
*Team 2: Seeded 8 colonies from ligation plate for miniprep next day.<br />
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'''July 14'''<br />
*Team 2: Miniprepped 8 seedings, using 5 mL of broth culture for each miniprep. Screened the minipreps using Not I and Sca I, expecting bands at 2kb, 1500 bp, and 500 bp. Incubated for a couple of hours. When gel ran, observed bands at 4 kb, and a wide band at 3500 kb - perhaps the plasmid was only cut once, and the remaining uncut plasmid caused the wide band.<br />
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'''July 16'''<br />
*Team 2: Set up screening digests for 4 of July 14 minipreps, again using Not I and Sca I in case the problem was due to lack of incubation time. Incubated overnight.<br />
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'''July 17'''<br />
*Team 2: Gel ran of last night's digests showed no DNA, and smeary ladder. Jay suggested our minipreps may not be clean enough, so we washed them with Buffer PB. New screening digests with same enzymes, ran gel and found that there was no ladder shown! Replaced TAE buffer - maybe it's eating our DNA. New screening digest set up for overnight incubation with Ase I.<br />
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'''July 18'''<br />
*Team 2: Ran gel of last night's digests, expected bands at 1920 bp and 2200 bp. Observed band at 3000 bp. Set up another screening digest of all 8 minipreps from July 14, using Not I and Eco RV. <br />
<br />
*Team 3: Team 3 is making its triumphant return after a two week hiatus from the lab notebook!!! We first prepared a concentrated miniprep of the psp-luc+ by using 250uL of Buffer P1 to resuspend all 5 cell pellets. This miniprep was then digested with EcoRi and Hind 3 (digest labeled "psp-luc+mini prep conc 7/18"). The digested miniprep is placed in Cloning Fragments 6D and 6E. The "conc mini psp-luc+ 7/14" was also digested. The psp-luc+ and pGFP which had been digested yesterday was run on a gel. The gel was successful and the GFP vector band was seen at 3300bp while the luciferase gene band was seen at 1700 bp. The vector and the gene DNA was then extracted from the gel. The extracted DNA, the GFP vector "GFP 7/18" was placed in 6G and the luc gene "luc gene 7/18 was placed in 6F :D<br />
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'''July 19'''<br />
*Team 3: The "luc mini 7/17" and "luc mini conc 7/14" digests were screened on gel, both yielding faint luc gene bands (1.7kb). The ligation of "luc gene 7/18" and "GFP (vector) 7/18" was performed: 15uL insert, 2uL vector, 2uL lig. buffer, 1 uL ligase. Incubated for 2 hrs at rm temp. 2 vials of Top10F' (1 commercial and 1 homemade) were transformed with the ligation, and plated.<br />
<br />
'''July 20'''<br />
*Team 3: Yesterday's ligation plates were removed from 37C and placed at 4C (at 11:00). The plate with homemade transf. Top10F' had no colonies, but the 2 plates with commercial transf. Top10F' had medium-sized colonies. To be seeded at 16:00. <br />
<br />
'''July 21'''<br />
*Team 3: A miniprep was prepared of the succesful seedings of the ligated GFP vector and the luciferase insert. The miniprep has now been digested to make sure that the DNA is in fact the ligated GFP and luciferase insert<br />
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'''July 25'''<br />
*Team 3: The digestion result revealed a few very important things. 2 minipreps out of 4 having been digested showed bands at 10kb and 8kb, and the other two at 4kb. The latter ones might contain the desired ligation product with the possibility of Sca1 inefficiency. The expected result from the digetion was 2300kb and 1900kb bands (adding up to approx 4kb, shockingly similar to the single bands that appeared on the gel). The minipreps of those latter ones are stored in the cloning fragment box (approx 30-50uL in each tube). A link to the paper about mechanotransduction activity in living cells has been posted under Journal Club Paper. Further analysis will be available after the group disccusion once Jieun snaps out of whatever she's going through.<br />
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'''July 25'''<br />
*Team 3: The 6 ligation minipreps (prepared yesterday) were digested with ScaI, and screened on gel. Again strange (very faint) bands appeared, this time above the DNA ladder. Possibilities are that these bands come from garbage DNA (contamination or chromosomal DNA), or ScaI isn't slicing properly! We should screen the minipreps to see if they actually any useful DNA, and try digesting with another enzyme.<br />
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'''July 27'''<br />
*Team 3: '''News To Horia:''' The Day Of Truth Has Arrived. According to Jieun "the truth is out there in the biohazard box." (long story- Jieun..was being herself = moody and Ashwini was being calm and rational) This time we digested with our best-buddies - EcoR1 and Hind III- and they did not dissappoint. The time had come, and the gel was placed on UV machine with Elvis' shaking hands. Jieun pressed the "Acquired" button, and voila. Did I say Voila? VOILA. BEAUTIFUL TWO SEPARATE BANDS PLACED WHERE THEY WERE SUPPOSED TO BE. We embraced this joyful moment by excessive hugging time and Jieun, despite her cranky and emotionally detached self, gave a big hug to her teammate,Ashwini. So, long story short. WE SUCCEEDED. BOOOOYAH. OHHHHH SNAPPPP!!!!<br />
<br />
'''July 28'''<br />
*Team 2: WORKING LIGATION!!<br />
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'''July 29'''<br />
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Notes from Jay:<br />
*Team 1: Adam worked till late hours purifying the gel pieces, but didn't ligate because the ligase was locked upstairs and we're out.<br />
Will anyone from Team 1 be in this weekend? I can do it if you like? Please email me if you're Team 1 and want to do something this weekend, I will be downtown this afternoon.<br />
To everyone, notes about the new Gel Purifcation Kit: please use ONLY ONE column for each piece that you cut out, no matter how much it weighs (try to minimize excess agarose). You can spin more than once. We only get 50 columns in the kit so please don't waste! Also, if you're purifying a piece with a molecular weight close to that of the loading dye (either the green or the purple), you will get some loading dye into your agarose and this will turn your melted piece greenish or purplish. This is OK and doesn't mean that you need to add sodium acetate. In fact, I have never once in many many many hundreds of these had to add sodium acetate. Always do the isopropanol addition, and the optional wash isn't necessary.<br />
C'est tout, bonne fin de semaine!<br />
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'''August 9'''<br />
*Team 3: To do the luciferase protein assay, we don't need to lyse the cells to react luciferase with luciferin. Jay found a procedure which can be found here: http://www.btci.org/k12/bft/bgt_background.html<br />
Our present goal is to make LAR (luciferase assay reagent - 1 mM luciferin in 100 mM sodium citrate pH 5.5). We will make the luciferin and sodium citrate solutions separately. The overall plan is to prepare a seeding (in LB) of our ligation cells with O.D. of ~0.3, induce it with IPTG, centrifuge, dissolve the pellet (cells) in LAR, and thence observe the expected bioluminescent reaction.<br />
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'''August 10'''<br />
*Team 3: 500 mL of 100 mM sodium citrate (pH 5.5) was made yesterday and store at 4C. Today, we diluted a seeding of pGFP-luc cells (made on Aug. 8 by Ashwin) 1:150 - 600 uL seeding in 30 mL fresh 1X LB with 30 uL. Placed in shaking incubator until O.D. measured to be 0.365. 30 uL IPTG added to cells (1 uL IPTG/1 mL total volume), and placed in shaker for 2 hours. Also found that luciferin can enter E. coli without need of MscL channel ...<br />
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'''August 23'''<br />
*Team 3: The new plates with pGFP-Luc had somewhat less of a lawn than the previous ones. A miniprep screening digest was performed by Ashwin - with similar results as the previous screening done by Alida. It seems the bands of DNA move more slowly than the ladder, and the bands are thick and almost but not exactly in the right place. Please refer to the gel photo. New seedings have been done from the new plate.<br />
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'''August 24'''<br />
*Team 3: Beetle Luciferin is scheduled to arrive tomorrow (Friday). A seeding of ligation cells - diluted in fresh LB at an O.D. of 0.3-0.4 and stimulated with IPTG - must be spun down. Discard the supernatant, and the pellet of cells must be resuspended in 3-5 mL of the prepared sodium citrate. Transfer 1-1.5 mL of this solution to a microcentrifuge tube, and add ~0.25 mg of luciferin. The bioluminescence must be viewed in a dark room. The quantities described above (determined by Elvis and Ashwin) are estimated and may not be totally accurate.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Registration_FAQRegistration FAQ2007-02-15T20:29:02Z<p>Nadeau: /* Structured Competition */</p>
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<div>[[image:buttonquestion.gif]]<br />
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== When does registration for iGEM 2007 start? ==<br />
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A registration page will be available later this summer. If you require additional information to prepare for the 2007 competition, please e-mail the organizers at igem@mit.edu<br />
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<br />
== What is iGEM about? ==<br />
<br />
<br />
'''A summary of the iGEM program can be found [http://icampus.mit.edu/projects/iGem.shtml here]'''.<br />
<br />
<br />
Basically, the iGEM competition and the Registry of Standard Biological Parts (aka 'the Registry')are practically testing the idea that biological engineering can be made reliable through the use of freely-shared, standardized, and well-documented parts called ''biobricks''. Biobricks have special features that allow for iterative assembly of longer "composite parts", leading to larger devices or systems.<br />
<br />
<br />
iGEM teams are challenged to create working devices by designing and assembling a) parts that already exist in the Registry and b) their own parts that conform to the biobrick standard. New parts must be documented in the Registry database and physically sent to the registry. This allows future biological engineers to build on the creativity, expertise, and experience of the people and groups that came before them.<br />
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== What are teams actually competing to do? == <br />
<br />
The goal of iGEM is currently to develop the "coolest" system. What your team chooses to do will depend a lot on what they perceive to be cool and also what they think they might actually be able to pull together. Innovation is encouraged. Jaw-dropping creativity, originality, and functionality will certainly be factors in the Judge's decisions of relatively coolness.<br />
<br />
There has been some discussion of adding another category to iGEM that would see teams compete towards the same goal (e.g. making the best functioning counter). <br />
<br />
<br />
== What is the iGEM schedule? == <br />
<br />
The program formally begins in May with the [[Teach the Teachers]] Workshop and closes with the [[Jamboree]] in November. The exact start and stop dates are variable depending on the team or institution requirements. Some teams begin to organize as early as March, while UK teams typically start as late as June.<br />
<br />
== What are the qualifications for entry? == <br />
<br />
Currently, there are two categories of competition.<br />
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=== Structured Competition ===<br />
<br />
The first category requires institution to assemble a formal iGEM team and to meet minimum participation standards. Aspects of the competition will be judged and prizes awarded. In order to be considered for the structured competition, teams must:<br />
<br />
* Be supervised by a qualified principal investigator<br />
* Be a mostly undergraduate team, although graduate students are permitted<br />
* Be based at an accredited college or university, with access to laboratory facilities<br />
* Send a representative to the iGEM Teachers Conference in May 6, 2006, at MIT. (EU date TBD)<br />
* Post regular updates of strategies and progress of their projects on the iGEM wiki<br />
* Freely share all materials developed for the competition (data, physical DNA)<br />
* Coordinate and support travel for team members to attend the iGEM [[Jamboree]] at MIT<br />
* Make a verbal presentation of length 20 minutes at the Jamboree<br />
* Deposit your project description on the iGEM Wiki and your parts in the Registry before the Jamboree<br />
* Prepare a poster presentation for the Jamboree<br />
* Have fun!<br />
<br />
=== Unstructured Competition, aka "Freestyle"===<br />
<br />
There are no rules or requirements, although you are welcome to contact the organizers if you have questions or concerns. Or not. You decide.<br />
<br />
== What does MIT provide iGEM teams? == <br />
<br />
MIT coordinates the iGEM competition, provides space and facilities for the conference and [[jamboree]], maintains tools and databases used to create projects, provides [[Biobrick delivery|Biobrick construct assemblies]] to teams, and organizes widespread publicity. In 2006, an ambassador program was established to provide additional support for teams.<br />
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== How should teams be structured? ==<br />
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We recommend a 50-50 guideline: 50% male, 50% female, 50% biological background, 50% non-biological, and so on. We now require that teams be primarily undergraduates, although graduate students are also permitted. At the institution's discretion, so are other students, including those may not be directly associated with your institution, like those from nearby high schools or community colleges. While iGEM student guidelines are not fixed in stone, we have learned that the experience is enriched by collecting a wide range of aptitudes, experiences, and skills for your team. We suggest advertising for team members, since this is likely to generate a large and diverse pool of applicants.<br />
<br />
== How much does it cost to participate? == <br />
<br />
This can vary widely by institution.<br />
<br />
Costs include:<br />
<br />
* salaries for instructors, if any<br />
* stipends to students, if any<br />
* reagents and supplies – variable depending on project<br />
* DNA synthesis costs - assume about $4000<br />
* travel support for the Teachers Conference and Jamboree – dependent on physical proximity to MIT<br />
* miscellaneous expenses<br />
<br />
== I am not affiliated with a university or college. Can I still participate? ==<br />
<br />
Yes. If you are located near a participating institution, you could contact the team organizer and apply for a position on the team. Even if the team cannot accept you as a full time member, they may still permit an internship or volunteership of some kind. Alternatively, you could participate in the unstructured competition, which has no rules. What you accomplish in this category is entirely up to you or your group. Finally, you can contact the organizers and tell them how you think you might be able to contribute.<br />
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<br />
== General Information to prepare for iGEM ==<br />
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== For Team Leaders ==<br />
<br />
* '''Announce:''' Announce your participation in iGEM early and widely with your peers. You are on the leading edge of an exciting event that is rapidly growing and gaining global recognition.<br />
<br />
* '''Advertise and recruit:''' Begin recruitment well before the spring term ends. Promote and advertise widely on campus. You are likely to get a surprising number of applications. Assembling a diverse team will bring new perspectives to the task of engineering biological systems.<br />
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* '''Undergraduates:''' Try to field a mostly undergraduate team. You will be surprised at how much they can do in so short a time. (They don't know what is impossible, yet!)<br />
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* '''Instructors:''' Find two instructors for the team. The summer is long and a single instructor may not be able to spend the whole summer supporting the team. Besides, it will be more fun. The instructors get as much from the summer as the students do.<br />
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* '''Workspace:''' Consider the workspace available to your team. A space large enough to permit them to gather and work together side by side will lead to a better team experience.<br />
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* '''Motivation:''' What is your motivation for participation? What do you expect from your team? Understanding what you hope to accomplish by competing in iGEM will help set the tone of the team you build.<br />
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* '''Teacher's Conference:''' Select a representative to attend the iGEM teachers conference in May. This is an important choice: this person will disseminate the information from the conference and is expected to act as a front line resource person for the team. Choose someone that has the free time to support the students and is considered approachable. See the [[Teach the Teachers]] page for more information.<br />
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* '''Wikis:''' Information sharing is central to iGEM, and this year we are requiring use of the iGEM wiki. Some teams also create their own wikis and external sites to augment the iGEM wiki, but the iGEM wiki is expected to be the main site for sharing information about each team's project and progress. <br />
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*'''What to Wiki:''' We encourage creative wiki-ing! However, we would like to distill meaningful summaries at the end of the competition. To facilitate this, we advise that the following information be included on each team's wiki pages:<br />
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# A list of all team members, their roles, and email addresses<br />
# Overview of project(s), including schematics and figures<br />
# Ongoing data/updates about project(s), including schematics, figures, test data, and biobrick parts used<br />
# Some photos of your team, facilities, institution, etc.<br />
# Optionally, anything that broadcasts your team's personality, spirit, sense of fun, or coolness...<br />
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* '''Parts registry:''' The [http://parts2.mit.edu/ parts registry] is a core tool of the iGEM program and worth the effort to experiment with sooner than later.<br />
'''Note:''' ''Development work is continuing on parts registry and a new version will launch before iGEM begins, complete with new features, data cleanup, and more complete documentation. This work will be completed before the teachers conference in May.''<br />
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* '''Project Planning:''' It's never too early to start thinking about what to make.<br />
<br />
* '''Journal Clubs:''' Excellent for getting the teams to think about technical issues and develop presentation skills. And getting pizzas delivered.<br />
<br />
* '''Fundraising:''' iGEM participation can require considerable financial support. Think about which organizations or groups might be willing to sponsor your team. Don't overlook the support of other departments at your institution. Make a list. When your team is assembled, consider writing letters requesting their help. You may want to have students host a talk outlining their projects and interests, which will help polish those presentation skills.<br />
<br />
* '''Media''' The media is interested in projects like iGEM that involve teams of young people and new, exciting technologies. iGEM is receiving more attention each year. Former projects are even appearing in peer reviewed articles in top journals. Media exposure will help teams take their work seriously, increase sponsorship and grant support, attract new talent, and more. You should consult with your institution's media office about contacting campus papers, local newspapers, and television.<br />
<br />
== For Students ==<br />
<br />
=== Why participate? ===<br />
<br />
iGEM is an opportunity to gain practical experience in collaborative scientific research in a casual yet supportive environment. A few of the benefits reported by former iGEMers include:<br />
<br />
* Chance to do self-directed, original research, not just menial summer work<br />
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* Experience in competitive science, but in a friendly and supportive environment<br />
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* Use of leading edge tools and methods towards practical applications<br />
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* Problem solving skill development<br />
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* Chance to interact with supervisors<br />
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* Opportunity to improve written and verbal presentation skills<br />
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* Inclusion in an international peer network<br />
<br />
=== Materials ===<br />
<br />
Check our [[Resources]] page to learn more about iGEM and Synthetic Biology in general.<br />
<br />
A lot of [http://openwetware.org/wiki/Synthetic_Biology:BioBricks useful information on BioBricks] is also available on OpenWetWare.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-11-04T12:55:31Z<p>Nadeau: </p>
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<h2>Fluorescence complementation</h2><br />
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<br />
<br />
<h4><font face="Broadway, Verdana">Conclusions</font></h4><br />
<br />
Fos Jun Complementation is indeed happening. However, the induction of bacteria does not last a long time, breaks at room temperature within a few hours. Also it is not quite sure whether the cells are touching while the complementation takes place or are simply close to each other.<br />
<br />
<h4><font face="Broadway, Verdana">Future Work</font></h4><br />
<br />
*Mutational nalysis must be performed to turn our plasmids into BioBricks and this includes mutations of Pst and other sites.<br />
*Work must be done to further understand the interesting formations we saw.<br />
*More inductions must be done to understand exactly the fluorescence pattern.<br />
*More understanding regarding stability of the protein products will be investigated.<br />
<br />
<h4><font face="Broadway, Verdana">Possible Future Work in the field</font></h4><br />
<br />
Fos and Jun complementation and half YFP can have a variety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins, eventually one could use bacteria as building blocks for entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-11-04T12:54:54Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Conclusions</font></h4><br />
<br />
Fos Jun Complementation is indeed happening. However, the induction of bacteria does not last a long time, breaks at room temperature within a few hours. Also it is not quite sure whether the cells are touching while the complementation takes place or are simply close to each other.<br />
<br />
<h4><font face="Broadway, Verdana">Possible Future Work</font></h4><br />
<br />
*Mutational nalysis must be performed to turn our plasmids into BioBricks and this includes mutations of Pst and other sites.<br />
*Work must be done to further understand the interesting formations we saw.<br />
*More inductions must be done to understand exactly the fluorescence pattern.<br />
*More understanding regarding stability of the protein products will be investigated.<br />
<br />
<h4><font face="Broadway, Verdana">Possible Future Work in the field</font></h4><br />
<br />
Fos and Jun complementation and half YFP can have a variety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins, eventually one could use bacteria as building blocks for entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-11-04T12:53:46Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Conclusions</font></h4><br />
<br />
Fos Jun Complementation is indeed happening. However, the induction of bacteria does not last a long time, breaks at room temperature within a few hours. Also it is not quite sure whether the cells are touching while the complementation takes place or are simply close to each other.<br />
<br />
<h4><font face="Broadway, Verdana">Possible Future Work</font></h4><br />
<br />
*Mutatuional nalysis must be performed to turn our plasmids into BioBricks and this includes mutations of Pst and other sites.<br />
*Work mustbe done to further understand the interesting formations we saw.<br />
*More inductions must be done to understand exactly the fluorescence patrtern.<br />
*More understanding regarding stability of the protein products is required.<br />
<br />
<h4><font face="Broadway, Verdana">Possible Future Work in the field</font></h4><br />
<br />
Fos and Jun complementation and half YFP can have a variety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins, eventually one could use bacteria as building blocks for entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-11-04T12:46:53Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Conclusions</font></h4><br />
<br />
Fos Jun Complementation is indeed happening. However, the induction of bacteria does not last a long time, breaks at room temperature within a few hours. Also it is not quite sure whether the cells are touching while the complementation takes place or are simply close to each other.<br />
<br />
<h4><font face="Broadway, Verdana">Possible Future Work</font></h4><br />
<br />
*Mutatuional nalysis must be performed to turn our plasmids into BioBricks and this includes mutations of Pst and other sites.<br />
*Work mustbe done to further understand the interesting formations we saw.<br />
*More inductions must be done to understand exactly the fluorescence patrtern.<br />
<br />
<h4><font face="Broadway, Verdana">Possible Future Work in the field</font></h4><br />
<br />
Fos and Jun complementation and half YFP can have a variety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins, eventually one could use bacteria as building blocks for entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-11-03T00:26:26Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Conclusions</font></h4><br />
<br />
Fos Jun Complementation is indeed happening. However, the induction of bacteria does not last a long time, breaks at room temperature within a few hours. Also it is not quite sure whether the cells are touching while the complementation takes place or are simply close to each other.<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Future Work</font></h4><br />
<br />
Fos and Jun complementation and half YFP can have a variety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins, eventually one could use bacteria as building blocks for entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-11-03T00:25:26Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Conclusions</font></h4><br />
<br />
Fos Jun Complementation is indeed happening. However, the induction of bacteria does not last a long time, breaks at room temperature within a few hours. Aslo it is not quite sure whether the cells are touching while the complementation tkes place or are simply close to each other.<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Future Work</font></h4><br />
<br />
Fos and Jun complementtion and half YFP can have a viiety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins,eentually one could use bacteria as building blocksfvo entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/More_DetailedMore Detailed2006-11-02T15:02:36Z<p>Nadeau: </p>
<hr />
<div>'''More detailed:'''<br />
<br />
'''The Vectors:'''<br />
<br />
*The vector plasmids Fos-Beta and Jun-Beta arrived from Spain on the 21st of June 2006. They arrived in the form of bacteria in agar tubes. This is from a Spanish group that bubplished the paper “Autotransporters as Scaffolds for Novel Bacterial Adhesins: Surface proteins of Esherishia Coli Cells displaying Jun/Fos Dimerization Domains” By Estaban Veiga, Víctor de Lorenzo, and Luis Angel Fernández. The plasmids had chloramphenicol resistance.<br />
<br />
*They have been seeded on June 28 at 11:45 am in 2ml 1X LB (without glucose) and 210ng of chloramphenicol (105ng/ml). This grew at 37OC until 20:00pm. At 20:00 pm 500μL of seedings were diluted to 25ml of 1X LB and 750ng chloramphenicol (30ng/ml). This grew overnight and at 9:56 am on June 29th a midiprep was performed using a quiagen midiprep kit. Each plasmid was dissolved in 50μL dH2O.<br />
<br />
*On July 4th the Jun-Beta plasmid midiprep DNA was confirmed by a restriction digest with BamHI that should have yielded two pieces at 1442bp and 3920bp. The digest was incubated at 37OC for 3 hours 23 minutes. This was ran on a 0.7% agarose gel with ethidium bromide and in comparison to known molecular weight markers, the fragments have been confirmed.<br />
<br />
*On July 6th the Fos-Beta DNA was confirmed by the exact same BamHI digestion at 37OC for 2h 20min, to yield pieces of similar size to the Jun-Beta, which it did.<br />
<br />
*On July 28th the concentrations of the midiprep DNAs were found using spectrophotometry to be 5950ng/μL for Jun-Beta and 4500ng/μL for Fos-Beta. This was done using 1000 fold dilution of original stock, a spectrophotometer set at 250nm and Quartz cuvettes.<br />
<br />
*On August 16th both the midipreps were diluted 5 fold with dH2O.<br />
<br />
*On September 12th a double digest was performed on both vectors with SacII and EcoRI. The SacII is an enzyme that takes longer t digest than EcoRI. Therefore digestion was first made with SacII in NEB buffer 4 using 5μL of DNA. These digests had a total volume of 50μL. They were incubated at 37OC for 3 hours. After about 3 hours 1μL of the digest was ran on a 0.7% agarose gel with ethidium bromide and in parallel lanes each uncut plasmid (point of having these lanes is to see cut compared to uncut, supercoiling). This indeed confirmed that the SacII hadentirely cut he Jun and almost entirely for fos.<br />
<br />
*To the sacII digests, ecoRI was added along with ecoRI buffer and diluted up to 100μL in a digest consisting of a half half mix of ecoRI buffer and buffer 4. This was incubated at 37OC for 1h.<br />
<br />
*Then the enzymes were deactivated by heating at 65OC for 20 minutes.<br />
<br />
*From the gel, by comparing to DNA ladder known marker DNA concentrations and using an optical system that quantitatively measures the intensity of the bands, the concentrations of the cut vectors were found to be 28.18ng/μL for Fos-Beta and 8.01ng/μL for Jun-Beta.<br />
<br />
<br />
<br />
'''Inserts:'''<br />
<br />
-PCR template:<br />
<br />
*On the 27th of June, what was to be the PCR template for both our inserts arrived from Michigan in the form of DNA. The plasmid was called pDsBiFc- bJunYN155- bFosYC155. This plasmid has Amp resistaqnce. It came from the team that wrote the article, "Visualization of Interactions among bZIP and Rel Family Proteins in Living Cells Using Bimolecular Fluorescence Complementation" by Chang-Deng Hu, Yurii Chinenov, and Tom K. Kerppola of Howerd Hughes Medical Institute and department of Biological CHemistry,<br />
<br />
*3μL of this DNA has immediately been transformed by heat shock into Top 10 chemically competent cells (One Shot). This was plated on two Ampicillin plates, 150μL, and 50μL transformant.<br />
<br />
*On June 28th, it was found that the plates had colonies.<br />
<br />
*On July 6th, 10 seedings from plates were done by 13:10 (in 2ml LB and Amp). They were incubating until 22:00 pm (8 hours 50 minutes) when they were re-suspended in QUIAGEN buffer P1 (by spinning down, removal of supernatant, adding 250μL buffer P1), and placed in fridge.<br />
<br />
*On July 10th, the re-suspended 9 of the seedings were re-seeded each in 5ml LB and Amp. They were placed at 37OC in shaker at 16:45pm.<br />
<br />
*On July 11th, The seedings were incubating very long until 15:30 pm (22h 45min) before a standard QUIAGEN miniprep procedure was performed on all of them.<br />
<br />
*On July 17th, first the miniprep DNA required purification using a QUIAGEN purification kit. Then, 8 double digests of purified DNA were se up at 18:35pm. They consisted of EcoRI, BamHI, NEB EcoRI buffer, and BSA. They were left at 37OC overnight.<br />
<br />
*The digests were run on a 0.8% agarose gel with ethidium bromide. The result of the gel is that the first of the eight digests had the expected band location, one at 4000bp and the other at 800bp.<br />
<br />
*To serve as a template, a 50X dilution of the first miniprep was performed.<br />
PCR primers:<br />
<br />
*Note: we are using two different sets of primers to PCR out two halves of YFP that are in the same plasmid pDsBiFc- bJunYN155- bFosYC155 (2 fusion proteins, 1 plasmid)<br />
<br />
*On July 5th, the primers were designed, ordered and arrived. Note: The melting temperatures were determined using software called Primer Express. So four primers, both forward primers had EcoRI site in the forward primer and SacII site in the back Primer<br />
<br />
*On July 7th, the dry primers were diluted with QUIAGEN buffer EB. (236μL EB Jun forward, 220μL EB Jun reverse, 170μL EB fos forward, 211μL EB Fos reverse.<br />
<br />
-PCR:<br />
<br />
*On July 27th, four PCR reactions were set up, two for each YFPN and YFPC (two halves of YFP). This was in a 100μL reaction vial each consisting of 80μL PCR dH2O, 10μL PCR buffer, 4μL dNTP’, 2μL each forward and reverse primer, 1μL BSA, 1μL Taq DNA Polymerase, 1μL Template (diluted pDsBiFc). Tese reaction tubes have undergone PCR. Following this the PCR solution was purified using PCR purification kit.<br />
<br />
*2.5μL of PCR products were run on a 1.2% agarose gel with ethidium bromide. On July 28th, using band comparison with ladder, the concentration of the PCR products was found to be about 25ng/μL (the intensity of all four bands was about the same).<br />
-Preparation of insert by digestion:<br />
<br />
*On August 23, a digestion of 20μL of the first of each PCR reactions was performed. EcoRI digestion was set up with NEB EcoRI buffer at 37OC for 3 hours. This was followed by running on a 1.2% agarose gel with ethidium bromide for about 15-30 minutes. Then a standard Gel extraction was performed using a Quiagen gel extraction kit (columns). This was followed by an overnight digestion with SacII in buffer 4 at 37OC. Then on August 24th, the digests were heat deactivated at 65OC for 20 minutes and 6μL of them was ran on a 1.2% agarose gel with EtBr and from this gel, the concentration of the inserts was found to be 14.6ng/μL for YFPC (Fos) and 8.95ng/μL for YFPN (Jun).<br />
<br />
'''Ligations'''<br />
<br />
*On September 13th, two ligations have been performed. One was a ligation of YFPC to Fos-Beta and the other a ligation of YFPN to Jun-Beta. It was decided, that both ligations were to include 100ng of insert and 100ng of vector. Knowing the concentrations of YFPC insert to be 14.6ng/μL, it was determined that 6.85μL would be used. Knowing Fos-beta concentration of 28.18ng/μL, 3.55μL would be used. In the other ligation, the concentrations were too small for 100ng of each to be used. Therefore, 59ng of each was used to keep the total volume under 20μl. Volume of YFPN was 6.61μL and Jun-Beta was 7.39μL. These ligations were left overnight in a room of about 18OC.<br />
<br />
*On September 14th, a transformation of the ligation product into homemade chemically competent Top 10 cells was performed and plated on chloramphenicol plates.<br />
<br />
*On September 15th, it was found colonies grew on the plates and there were not too many or too few.<br />
<br />
*On September 21st, a protocol referred to as “cracking” was performed. Several colonies were picked from each ligation, they were placed in 20μL sterile dH2O, mixed and 1μL was placed on another plate with labelled grid. This was heated at 95OC for 5 minutes and then PCR could be performed directly on these, so several PCR reactions were performed.<br />
<br />
*On September 22nd, five of the YFPN (Jun) PCR products were loaded onto a 1.2% gel. Bands of the right sizes were found (486bp) and that was the case for four out of the five wells had the right bands. On September 25th, nine of the YFPC (Fos) PCR products ran on a gel and all resulted in the correct band at 281bp.<br />
<br />
'''iPTG induction and viewing'''<br />
<br />
*On September 26th, 2mL LB seedings with 2ng/ml chloramphenicol of colonies two of each ligation were performed and placed in 37OC Shaker. Later that day a standard miniprep procedure was ran and on September 27th, a transformation into commercial Bl21 cells has been done. On September 28th, two colonies of each transformant were seeded in 4mL LB overnight. On September 29th, iPTG induction began: diluted 500μL of each seeding into a 5ml LB and chloramphenicol seeding for each, as well as another dilution of 1mL into 5mL. This grew in 37OC shaker and after 1 hour the OD measurement (600nm) was 0.318 for Jun 1L and 0.248 of Fos 500μL. They were induced with the addition of 0.3μL iPTG. They were grown for 1 hour and were spun down, dissolved in water (LB is fluorescent) and viewed under the microscope. The result appeared that when added together, sells did clump together and glow yellow. Therefore, this was a success. However, unfortunately the induced cells did not last over the weekend in the fridge in LB.<br />
<br />
*This success was repeated once more with another transformation. This time, on Wednesday October 25th, the iPTG induction was 3 hours and the cells were not spinned in centrifuge, instead they have undergone sedimentation at 1G. Some of them were mixed and incubated together while mixed. Again, a few hours later this induction did not last.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-10-31T20:24:03Z<p>Nadeau: </p>
<hr />
<div>'''Conclusions'''<br />
<br />
Fos Jun Complementation is indeed happening. However, the induction of bacteria does not last a long time, breaks at room temperature within a few hours. Aslo it is not quite sure whether the cells are touching while the complementation tkes place or are simply close to each other.<br />
<br />
'''Future Work'''<br />
<br />
Fos and Jun complementtion and half YFP can have a viiety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins,eentually one could use bacteria as building blocksfvo entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-10-31T20:23:48Z<p>Nadeau: </p>
<hr />
<div>'''Conclusions''<br />
Fos Jun Complementation is indeed happening. However, the induction of bacteria does not last a long time, breaks at room temperature within a few hours. Aslo it is not quite sure whether the cells are touching while the complementation tkes place or are simply close to each other.<br />
<br />
'''Future Work'''<br />
Fos and Jun complementtion and half YFP can have a viiety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins,eentually one could use bacteria as building blocksfvo entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-31T20:19:19Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Good_2.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
[[Future Prospects|Conclusions and Future Work]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
Our project is based on the repressilator system coupled to quorum sensing as described by Jordi Garcia-Ojalvo, Michael B. Elowitz and Steven H. Strogatz in "Modeling a synthetic multicellular clock: Repressilators coupled by quorum sensing" (PNAS). We attempt to visualize the synchronization of the oscillatory phase between cells by the addition of the CFP reporter gene. We expand on this theory by placing cI under the control of pLac, hoping that this would assist in synchronizing the oscillations.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-31T20:18:44Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Good_2.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
[[Conclusions and Future Work|Future Prospects]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
Our project is based on the repressilator system coupled to quorum sensing as described by Jordi Garcia-Ojalvo, Michael B. Elowitz and Steven H. Strogatz in "Modeling a synthetic multicellular clock: Repressilators coupled by quorum sensing" (PNAS). We attempt to visualize the synchronization of the oscillatory phase between cells by the addition of the CFP reporter gene. We expand on this theory by placing cI under the control of pLac, hoping that this would assist in synchronizing the oscillations.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/Future_ProspectsFuture Prospects2006-10-31T20:10:40Z<p>Nadeau: </p>
<hr />
<div>'''Future Prospects'''<br />
<br />
Fos and Jun complementtion and half YFP can have a viiety of applications. One that we were thinking about was to fuse either Fos or Jun with an element of the flagella and that would stick to the membrane of a next cell, hence forming a chain. By fusing to different membrane proteins,eentually one could use bacteria as building blocksfvo entire strucures that work in synchrony. What still has to be worked out is their short life span.</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-31T20:02:36Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Good_2.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
[[Future Prospects|Future Prospects]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
Our project is based on the repressilator system coupled to quorum sensing as described by Jordi Garcia-Ojalvo, Michael B. Elowitz and Steven H. Strogatz in "Modeling a synthetic multicellular clock: Repressilators coupled by quorum sensing" (PNAS). We attempt to visualize the synchronization of the oscillatory phase between cells by the addition of the CFP reporter gene. We expand on this theory by placing cI under the control of pLac, hoping that this would assist in synchronizing the oscillations.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-30T19:21:26Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Good_2.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
Our project is based on the repressilator system coupled to quorum sensing as described by Jordi Garcia-Ojalvo, Michael B. Elowitz and Steven H. Strogatz in "Modeling a synthetic multicellular clock: Repressilators coupled by quorum sensing" (PNAS). We attempt to visualize the synchronization of the oscillatory phase between cells by the addition of the CFP reporter gene. We expand on this theory by placing cI under the control of pLac, hoping that this would assist in synchronizing the oscillations.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-30T19:20:48Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Good_2.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
Our project is based on the repressilator system coupled to quorum sensing as described by Jordi Garcia-Ojalvo, Michael B. Elowitz and Steven H. Strogatz in "Modeling a synthetic multicellular clock: Repressilators coupled by quorum sensing" (PNAS). We attempt to visualize the synchronization of the oscillatory phase between cells by the addition of the CFP reporter gene. We expand on this theory by placing cI under the control of pTet, hoping that this would assist in synchronizing the oscillations.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-30T18:56:18Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Good_2.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
Our project concerns the development of a repressilator coupled to quorum sensing as described by Jordi Garcia-Ojalvo, Michael B. Elowitz and Steven H. Strogatz in PNAS. The Elowitz Repressilator attempts to decrease the loss of standard oscillations that previous repressilators faced by utilizing quorum sensing as a means of synchronizing and maintaining standard oscillations. We expanded on this theory by adding YFP and CFP to allow a visual confirmation of the oscillation, and a TetR promoter in front of the LuxR gene and cI after the LuxI gene. Our hopes were that this would assist in standardizing the oscillation of the bacteria.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/File:Good_2.jpgFile:Good 2.jpg2006-10-30T18:55:33Z<p>Nadeau: </p>
<hr />
<div></div>Nadeauhttp://2006.igem.org/wiki/index.php/File:Dancing_e_copy-final.jpgFile:Dancing e copy-final.jpg2006-10-30T18:45:22Z<p>Nadeau: </p>
<hr />
<div></div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-30T18:29:33Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Dancing e copyzz.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
Our project concerns the development of a repressilator coupled to quorum sensing as described by Jordi Garcia-Ojalvo, Michael B. Elowitz and Steven H. Strogatz in PNAS. The Elowitz Repressilator attempts to decrease the loss of standard oscillations that previous repressilators faced by utilizing quorum sensing as a means of synchronizing and maintaining standard oscillations. We expanded on this theory by adding YFP and CFP to allow a visual confirmation of the oscillation, and a TetR promoter in front of the LuxR gene and cI after the LuxI gene. Our hopes were that this would assist in standardizing the oscillation of the bacteria.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/BackgroundBackground2006-10-29T17:49:21Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h3><font face="Broadway, Verdana">Introduction</font></h3><br />
<br />
The idea behind the project is fluorescence complementation, which involves the joining of two leucine zipper proteins, Fos and Jun, each fused to a half terminus of YFP. Originally, the Fos and Jun proteins were fused to a beta gene coding for a membrane protein. The project involved performing a PCR reaction to produce two inserts, the N-terminus and the C-terminus of YFP, and then ligating these inserts into 2 vectors, containing Jun-beta and the Fos-beta respectively. The two fusion proteins (Fos-beta-YFPC and Jun-beta-YFPN) were expressed in the cell membrane of two populations of <i>E. coli</i>. We then allowed these two cell types to combine, resulting—ideally—in the complementary binding of the Jun and Fos proteins when the cells are in close contact. Consequently, the two half YFP fragments bind to form full YFP, and the cells will fluoresce.<br />
<br />
<br />
<br />
<br />
<center>[[Image:Intro1.JPG]] </center><br />
<br />
<br />
<h3>[[McGill_University_2006|Back to McGill University Main Page]]</h3></div>Nadeauhttp://2006.igem.org/wiki/index.php/IGEM_Party_PicturesIGEM Party Pictures2006-10-29T00:46:56Z<p>Nadeau: </p>
<hr />
<div>[[McGill_University_2006|Home]]<br />
<br />
<br />
[[Image:igem1.jpg|frame|left|Adam, Jieun, Ashwini, Brock, Ashwin, Horia]]<br />
[[Image:igem2.jpg|frame|left|Same + Aaron in the back]]<br />
[[Image:igem3.jpg|frame|left|iGEM on the ceiling :)]]<br />
[[Image:igem4.jpg|frame|left|Ly & Horia]]<br />
[[Image:igem5.jpg|frame|left|Ly & Ada. Alex in the back]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/Methods_and_MaterialsMethods and Materials2006-10-29T00:45:20Z<p>Nadeau: </p>
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| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
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<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h3><font face="Broadway, Verdana">Methods and Materials</font></h3><br />
<br />
<br />
'''This section constitutes the procedural aspect of Team 1'''<br />
<br />
*[[Basic Summary|Basic Summary]]<br />
<br />
*[[More Detailed|More Detailed]]<br />
<br />
<br />
<h3>[[McGill_University_2006|Back to McGill's Main Page]]</h3></div>Nadeauhttp://2006.igem.org/wiki/index.php/Methods_and_MaterialsMethods and Materials2006-10-29T00:42:26Z<p>Nadeau: </p>
<hr />
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| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h3><font face="Broadway, Verdana">Methods and Materials</font></h3><br />
<br />
<br />
'''This section constitutes the procedural aspect of Team 1'''<br />
<br />
*[[Basic Summary|Basic Summary]]<br />
<br />
*[[More Detailed|More Detailed]]<br />
<br />
<br />
[[McGill_University_2006|Back to McGill's Main Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-29T00:38:45Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Dancing e copyzz.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project on fluorescence complementation involves the joining of two proteins, Jun and Fos, each fused to a half terminus of YFP. Both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Then, two cell populations - one expressing Jun-beta-YFPN and the other Fos-beta-YFPC - were combined, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. The two halves of the YFP protein would bind as well, giving rise to fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
The Elowitz Repressilator attempts to decrease the loss of standard oscillations that previous repressilators faced by utilizing quorum sensing as a means of synchronizing and maintaining standard oscillations. We expanded on this theory by adding YFP and CFP to allow a visual confirmation of the oscillation, and a TetR promoter in front of the LuxR gene and cI after the LuxI gene. Our hopes were that this would assist in standardizing the oscillation of the bacteria.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-29T00:30:34Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Dancing e copyzz.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project involves the joining of two proteins Jun and Fos each fused to a half terminus of YFP. <br />
Subsequently, both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. Two cell populations - one expressing Jun-beta-YFPN and the other, Fos-beta-YFPC - were combined and the two proteins were expressed, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. This would result in the binding of the two halves of the YFP protein resulting in fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
The Elowitz Repressilator attempts to decrease the loss of standard oscillations that previous repressilators faced by utilizing quorum sensing as a means of synchronizing and maintaining standard oscillations. We expanded on this theory by adding YFP and CFP to allow a visual confirmation of the oscillation, and a TetR promoter in front of the LuxR gene and cI after the LuxI gene. Our hopes were that this would assist in standardizing the oscillation of the bacteria.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-29T00:29:12Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
<br />
[[Image:Clip_image002.jpg]][[Image:Dancing e copyzz.jpg]]<br />
<br />
<br />
<br />
<!--{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
|width=179.25px style="padding: 5px; background-color: #fffaf0; border: 2px solid #20b2aa;" |<br />
EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
<br />
<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
<br />
<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
This project involves the joining of two proteins Jun and Fos each fused to a half terminus of YFP. <br />
Subsequently, both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. The<br />
N- and C-terminus of YFP were obtained by PCR, then ligated into 2 vectors, containing Jun-beta and the Fos-beta respectively.<br />
Two cell populations - one expressing Jun-beta-YFPN and the other, Fos-beta-YFPC - were combined and the two proteins were expressed, ideally resulting in the fusion of the Jun and Fos proteins on the cell membrane. This would result in the binding of the two halves of the YFP protein resulting in fluorescence.<br />
<br />
[[Background|Background]]<br />
<br />
[[Methods and Materials|Methods and Materials]]<br />
<br />
[[Results|Results]]<br />
<br />
</td><br />
<br />
<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
<br />
The Elowitz Repressilator attempts to decrease the loss of standard oscillations that previous repressilators faced by utilizing quorum sensing as a means of synchronizing and maintaining standard oscillations. We expanded on this theory by adding YFP and CFP to allow a visual confirmation of the oscillation, and a TetR promoter in front of the LuxR gene and cI after the LuxI gene. Our hopes were that this would assist in standardizing the oscillation of the bacteria.<br />
<br />
[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
<br />
[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
<br />
[[McGill_Repressilator_Results|Results]]<br />
</td><br />
<br />
</tr></table><br />
<br />
<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
<br />
{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0px; width: 750px; color: #000000; background-color: #ffffff;"<br />
|-valign="top"<br />
<br />
<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
<br />
<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
<br />
__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:21:13Z<p>Nadeau: </p>
<hr />
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| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG|frame|left|<b>Figure 1.</b> Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.]]<br />
<br />
<br />
[[Image:Results2.JPG|frame|left|<b>Figure 2.</b> Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.]]<br />
<br />
<br />
[[Image:Results3.JPG|frame|left|<b>Figure 3.</b> Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.]]<br />
<br />
<br />
[[Image:Results4.JPG|frame|left|<b>Figure 4.</b> Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.]]<br />
<br />
<br style="clear:both;"/><br />
<br />
<br />
<h3>[[McGill_University_2006|Back to McGill University Main Page]]</h3></div>Nadeauhttp://2006.igem.org/wiki/index.php/BackgroundBackground2006-10-29T00:18:11Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h3><font face="Broadway, Verdana">Introduction</font></h3><br />
<br />
The idea behind the project is fluorescence complementation, which involves the joining of two leucine zipper proteins, Fos and Jun, each fused to a half terminus of YFP. Originally, the Fos and Jun proteins were fused to a beta gene coding for a membrane protein. The project involved performing a PCR reaction to produce two inserts, the N-terminus and the C-terminus of the YFP, and then ligating these inserts into 2 vectors, containing Jun-beta and the Fos-beta respectively. The two fusion proteins (Fos-beta-YFPC and Jun-beta-YFPN) were expressed in the cell membrane of two populations of E. coli. We then allowed these two cell types to combine, resulting—ideally—in the complementary binding of the Jun and Fos proteins when the cells are in close contact. Consequently, the two half YFP fragments bind to form full YFP, and the cells will fluoresce.<br />
<br />
<br />
<br />
<br />
<center>[[Image:Intro1.JPG]] </center><br />
<br />
<br />
<h3>[[McGill_University_2006|Back to McGill University Main Page]]</h3></div>Nadeauhttp://2006.igem.org/wiki/index.php/BackgroundBackground2006-10-29T00:17:44Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
== Fluorescence complementation ==<br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h3><font face="Broadway, Verdana">Introduction</font></h3><br />
<br />
The idea behind the project is fluorescence complementation, which involves the joining of two leucine zipper proteins, Fos and Jun, each fused to a half terminus of YFP. Originally, the Fos and Jun proteins were fused to a beta gene coding for a membrane protein. The project involved performing a PCR reaction to produce two inserts, the N-terminus and the C-terminus of the YFP, and then ligating these inserts into 2 vectors, containing Jun-beta and the Fos-beta respectively. The two fusion proteins (Fos-beta-YFPC and Jun-beta-YFPN) were expressed in the cell membrane of two populations of E. coli. We then allowed these two cell types to combine, resulting—ideally—in the complementary binding of the Jun and Fos proteins when the cells are in close contact. Consequently, the two half YFP fragments bind to form full YFP, and the cells will fluoresce.<br />
<br />
<br />
<br />
<br />
<center>[[Image:Intro1.JPG]] </center><br />
<br />
<br />
<h3>[[McGill_University_2006|Back to McGill University Main Page]]</h3></div>Nadeauhttp://2006.igem.org/wiki/index.php/BackgroundBackground2006-10-29T00:16:25Z<p>Nadeau: </p>
<hr />
<div>{| cellspacing="2px" cellpadding="0" border="0" style="padding: 0; width: 750px; color: #000000; background-color: #ffffff;"<br />
| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h3><font face="Broadway, Verdana">Introduction</font></h3><br />
<br />
The idea behind the project is fluorescence complementation, which involves the joining of two leucine zipper proteins, Fos and Jun, each fused to a half terminus of YFP. Originally, the Fos and Jun proteins were fused to a beta gene coding for a membrane protein. The project involved performing a PCR reaction to produce two inserts, the N-terminus and the C-terminus of the YFP, and then ligating these inserts into 2 vectors, containing Jun-beta and the Fos-beta respectively. The two fusion proteins (Fos-beta-YFPC and Jun-beta-YFPN) were expressed in the cell membrane of two populations of E. coli. We then allowed these two cell types to combine, resulting—ideally—in the complementary binding of the Jun and Fos proteins when the cells are in close contact. Consequently, the two half YFP fragments bind to form full YFP, and the cells will fluoresce.<br />
<br />
<br />
<br />
<br />
<center>[[Image:Intro1.JPG]] </center><br />
<br />
<br />
<h3>[[McGill_University_2006|Back to McGill University Main Page]]</h3></div>Nadeauhttp://2006.igem.org/wiki/index.php/BackgroundBackground2006-10-29T00:14:31Z<p>Nadeau: </p>
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<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
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<br />
<h4><font face="Broadway, Verdana">Introduction</font></h4><br />
<br />
The idea behind the project is fluorescence complementation, which involves the joining of two leucine zipper proteins, Fos and Jun, each fused to a half terminus of YFP. Originally, the Fos and Jun proteins were fused to a beta gene coding for a membrane protein. The project involved performing a PCR reaction to produce two inserts, the N-terminus and the C-terminus of the YFP, and then ligating these inserts into 2 vectors, containing Jun-beta and the Fos-beta respectively. The two fusion proteins (Fos-beta-YFPC and Jun-beta-YFPN) were expressed in the cell membrane of two populations of E. coli. We then allowed these two cell types to combine, resulting—ideally—in the complementary binding of the Jun and Fos proteins when the cells are in close contact. Consequently, the two half YFP fragments bind to form full YFP, and the cells will fluoresce.<br />
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<center>[[Image:Intro1.JPG]] </center><br />
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[[McGill_University_2006|Back to McGill University Main Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/BackgroundBackground2006-10-29T00:14:20Z<p>Nadeau: </p>
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<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<h4><font face="Broadway, Verdana">Introduction</font></h4><br />
<br />
The idea behind the project is fluorescence complementation, which involves the joining of two leucine zipper proteins, Fos and Jun, each fused to a half terminus of YFP. Originally, the Fos and Jun proteins were fused to a beta gene coding for a membrane protein. The project involved performing a PCR reaction to produce two inserts, the N-terminus and the C-terminus of the YFP, and then ligating these inserts into 2 vectors, containing Jun-beta and the Fos-beta respectively. The two fusion proteins (Fos-beta-YFPC and Jun-beta-YFPN) were expressed in the cell membrane of two populations of E. coli. We then allowed these two cell types to combine, resulting—ideally—in the complementary binding of the Jun and Fos proteins when the cells are in close contact. Consequently, the two half YFP fragments bind to form full YFP, and the cells will fluoresce.<br />
<br />
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<center>[[Image:Intro1.JPG]] </center><br />
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[[McGill_University_2006|Back to McGill University Main Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:13:58Z<p>Nadeau: </p>
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<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
[[McGill_University_2006|Back to McGill University Main Page]]<br />
<br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG|frame|left|<b>Figure 1.</b> Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.]]<br />
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<br />
[[Image:Results2.JPG|frame|left|<b>Figure 2.</b> Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.]]<br />
<br />
<br />
[[Image:Results3.JPG|frame|left|<b>Figure 3.</b> Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.]]<br />
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<br />
[[Image:Results4.JPG|frame|left|<b>Figure 4.</b> Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.]]<br />
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<br style="clear:both;"/><br />
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[[McGill_University_2006|Back to McGill University Main Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:12:56Z<p>Nadeau: </p>
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<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG|frame|left|<b>Figure 1.</b> Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.]]<br />
<br />
<br />
[[Image:Results2.JPG|frame|left|<b>Figure 2.</b> Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.]]<br />
<br />
<br />
[[Image:Results3.JPG|frame|left|<b>Figure 3.</b> Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.]]<br />
<br />
<br />
[[Image:Results4.JPG|frame|left|<b>Figure 4.</b> Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.]]<br />
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<br style="clear:both;"/><br />
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[[McGill_University_2006|McGill University Main Wiki Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/McGill_University_2006McGill University 20062006-10-29T00:10:29Z<p>Nadeau: </p>
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'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
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[[Image:Clip_image002.jpg]][[Image:Dancing e copyzz.jpg]]<br />
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EDITED BY ASHWIN. If you want to change it, feel free :)--><br />
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<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
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<table cellpadding="5" border="0"><tr><br />
<td valign="top" width="50%"><br />
<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
The idea behind the project is fluorescence complementation, which involves the joining of two leucine zipper proteins Jun and Fos each fused to a half terminus of YFP. <br />
Subsequently, both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. The project involved performing a PCR reaction to produce two inserts, the<br />
N-terminus and the C-terminus of the YFP, and then ligating these inserts into 2 vectors, containing the<br />
Jun-beta and the Fos-beta respectively.<br />
We transformed two cell populations, one expressing Jun-beta-YFPN and the other expressing Fos-beta-YFPC.These two cell populations were combined and the two vectors were then expressed, ideally resulting in the fusion of the Jun and Fos leucine zipper proteins on the cell membrane when the cells are in close contact. This would result in the binding of the two halves of the YFP protein resulting in flourescence.<br />
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[[Background|Background]]<br />
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[[Methods and Materials|Methods and Materials]]<br />
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[[Results|Results]]<br />
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</td><br />
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<td valign="top" width="50%"><br />
<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
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The Elowitz Repressilator attempts to decrease the loss of standard oscillations that previous repressilators faced by utilizing quorum sensing as a means of synchronizing and maintaining standard oscillations. We expanded on this theory by adding YFP and CFP to allow a visual confirmation of the oscillation, and a TetR promoter in front of the LuxR gene and cI after the LuxI gene. Our hopes were that this would assist in standardizing the oscillation of the bacteria.<br />
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[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
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[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
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[[McGill_Repressilator_Results|Results]]<br />
</td><br />
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</tr></table><br />
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<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
<table border="0"><tr><td valign="top" width="85%"><br />
* [[Protocols]]<br />
* [[Lab Notebook]]<br />
</td><br />
<td>[[Image:Test_tubes.jpg]]</td></tr></table><br />
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<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
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<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
<Center>[[Image:Poutine.gif]]</Center><br />
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__NOTOC__</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:09:51Z<p>Nadeau: </p>
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<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG|frame|left|<b>Figure 1.</b> Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.]]<br />
<br />
<br />
[[Image:Results2.JPG|frame|left|<b>Figure 2.</b> Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.]]<br />
<br />
<br />
<br />
<br />
[[Image:Results3.JPG|frame|left|<b>Figure 3.</b> Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.]]<br />
<br />
<br />
<br />
<br />
[[Image:Results4.JPG|frame|left|<b>Figure 4.</b> Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.]]<br />
<br />
<br style="clear:both;"/><br />
<br />
<br />
[[McGill_University_2006|McGill University Main Wiki Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:05:00Z<p>Nadeau: </p>
<hr />
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<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG|frame|left|<b>Figure 1.</b> Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.]]<br />
<br />
<br />
<br />
[[Image:Results2.JPG|frame|left|<b>Figure 2.</b> Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.]]<br />
<br />
<br />
<br />
<br />
[[Image:Results3.JPG|frame|left|<b>Figure 3.</b> Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.]]<br />
<br />
<br />
<br />
<br />
[[Image:Results4.JPG|frame|left|<b>Figure 4.</b> Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.]]<br />
<br />
<br />
<br />
[[McGill_University_2006|McGill University Main Wiki Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:04:18Z<p>Nadeau: </p>
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<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG|frame|left|Figure 1. Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.]]<br />
<br />
<br />
<br />
[[Image:Results2.JPG|frame|left|Figure 2. Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.]]<br />
<br />
<br />
<br />
<br />
[[Image:Results3.JPG|frame|left|Figure 3. Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.]]<br />
<br />
<br />
<br />
<br />
[[Image:Results4.JPG|frame|left|Figure 4. Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.]]<br />
<br />
<br />
<br />
[[McGill_University_2006|McGill University Main Wiki Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:03:33Z<p>Nadeau: </p>
<hr />
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<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG|frame|left|Figure 1. Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.]]<br />
<br />
<br />
<br />
[[Image:Results2.JPG|frame|left|Figure 2. Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.]]<br />
<br />
<br />
<br />
[[Image:Results3.JPG|frame|left|Figure 3. Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.]]<br />
<br />
<br />
<br />
[[Image:Results4.JPG|frame|left|Figure 4. Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.]]<br />
<br />
<br />
<br />
[[McGill_University_2006|McGill University Main Wiki Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:02:53Z<p>Nadeau: </p>
<hr />
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<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG|frame|Figure 1. Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.]]<br />
<br />
<br />
<br />
[[Image:Results2.JPG|frame|Figure 2. Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.]]<br />
<br />
<br />
<br />
[[Image:Results3.JPG|frame|Figure 3. Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.]]<br />
<br />
<br />
<br />
[[Image:Results4.JPG|frame|Figure 4. Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.]]<br />
<br />
<br />
<br />
[[McGill_University_2006|McGill University Main Wiki Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-29T00:00:10Z<p>Nadeau: </p>
<hr />
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| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
<br />
<br />
[[Image:Results1.JPG]]<br />
<br />
Figure 1. Mixing of Jun-YFP and Fos-YFP expressing cells. Cells were taken up into patch pipettes pulled to a diameter of 1 micron and broken against the culture dish.<br />
<br />
<br />
[[Image:Results2.JPG]]<br />
<br />
Figure 2. Mixing of Jun- and Fos-expressing cells in a culture dish using a patch pipette. The phase contrast image shows a concentrated solution of Jun-expressing cells alone; the middle panel shows the fluorescence from these cells. In the right-hand panel, Fos was added from gentle pressure against the patch pipette approximately 60 s before the image was taken.<br />
<br />
<br />
[[Image:Results3.JPG]]<br />
<br />
Figure 3. Cell clustering and pattern of fluorescence. Sometimes, but not always, mixing of the Fos- and Jun-expressing cells leads to the formation of cell clusters. In the right-hand panel, a fluorescence image of one of the clusters shows a speckled fluorescence pattern.<br />
<br />
<br />
[[Image:Results4.JPG]]<br />
<br />
Figure 4. Sometimes the fluorescence pattern appears more continuous and over the entire cell. Cells are usually paired but occasionally appear singly.<br />
<br />
<br />
<br />
[[McGill_University_2006|McGill University Main Wiki Page]]</div>Nadeauhttp://2006.igem.org/wiki/index.php/ResultsResults2006-10-28T23:58:53Z<p>Nadeau: </p>
<hr />
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| width=717px style="padding: 5px; background-color: #ffffff; border: 2px solid #006666;" |<br />
<center>[[Image:Banner.gif]]</center><br />
<br />
<br />
<h2>Fluorescence complementation</h2><br />
<br />
<h4><font face="Broadway, Verdana">Results</font></h4><br />
<br />
<br />
We think we see fluorescence complementation! However, the expression seems rather fragile. Leaving the tubes overnight in the fridge or pipetting bacteria onto a dry slide wipes out the expression. It’s also not completely clear if the pattern of expression makes sense—we expected to see cells fluoresce only where they touch. So there are still some details to work out, but we saw some cool things!<br />
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'''Welcome to the McGill Wiki!''' <br />
Basically, we're a team of 12 undergrads, 1 grad student, and 1 professor who like to have fun and clone things in our spare time. McGill University is located in Montreal, Quebec, which has given us wonderful opportunities to balance lab work with festivals and general craziness. iGEM has been a great opportunity to get lab experience while having the freedom to be creative, and we look forward to meeting the other teams at the jamboree!<br />
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<center><h2><font face="broadway,verdana">Projects</font></h2></center><br />
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<h3><ul><li>'''Team 1: Split YFP'''</ul></h3><br />
The idea behind the project is fluorescence complementation, which involves the joining of two leucine zipper proteins Jun and Fos each fused to a half terminus of YFP. <br />
Subsequently, both of these chimeric proteins were fused to a beta gene that codes for a membrane protein. The project involved performing a PCR reaction to produce two inserts, the<br />
N-terminus and the C-terminus of the YFP, and then ligating these inserts into 2 vectors, containing the<br />
Jun-beta and the Fos-beta respectively.<br />
We transformed two cell populations, one expressing Jun-beta-YFPN and the other expressing Fos-beta-YFPC.These two cell populations were combined and the two vectors were then expressed, ideally resulting in the fusion of the Jun and Fos leucine zipper proteins on the cell membrane when the cells are in close contact. This would result in the binding of the two halves of the YFP protein resulting in flourescence.<br />
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[[Background|Background]]<br />
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[[Methods and Materials|Methods and Materials]]<br />
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<h3><ul><li>''' Team 2: Repressilator'''</ul></h3><br />
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The Elowitz Repressilator attempts to decrease the loss of standard oscillations that previous repressilators faced by utilizing quorum sensing as a means of synchronizing and maintaining standard oscillations. We expanded on this theory by adding YFP and CFP to allow a visual confirmation of the oscillation, and a TetR promoter in front of the LuxR gene and cI after the LuxI gene. Our hopes were that this would assist in standardizing the oscillation of the bacteria.<br />
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[[Theory Behind the Oscillator|Theory Behind the Oscillator]]<br />
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[[McGill_Repressilator_M_and_M|Methods and Materials]]<br />
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[[McGill_Repressilator_Results|Results]]<br />
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<center><h2><font face="broadway,verdana">Lab Procedures</font></h2></center><br />
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* [[Protocols]]<br />
* [[Lab Notebook]]<br />
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<center><h2><font face="broadway,verdana">Club</font></h2></center><br />
* [[News]]<br />
* [[Team Members]]<br />
* [[Executive Council|Executive Council]]<br />
* [[Journal Club Meetings]]<br />
* [[Journal Club Papers]]<br />
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<center><h2><font face="broadway,verdana">Just for Fun</font></h2></center><br />
* [[iGEM Party Pictures]]<br />
* [[ahessel:montreal|Ambassador Visit]] [http://www.flickr.com/search/?q=igem+montreal&m=text Photos]<br />
* [[iGEM Soundtrack]]<br />
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