Hometown: East Lyme, CT
I helped to divide our new idea of the bacterial freeze tag into four smaller projects. Jamie Lemon and I began working on the project called the freeze machine. We successfully transformed all of the parts required for this project. We also began miniprepping our parts, but were unable to finish due to a shortage of miniprep columns. We will finish the minipreps next week and then proceed onto digestion and ligations.
Wednesday: Performed transformations for Freeze machine parts. Made Amp. plates. Thursday: Updated wiki with Jamie Lemon and made of list of tasks for the group. We also grew up colonies for a miniprep tomorrow.
Friday: The colonies were not placed in the correct incubator and they therefore did not grow properly. I placed the colonies in the incubator during the day and Azeema and Jason will do the miniprep tonight. I also retransformed the colonies B0033 and C0260.
Sunday: I came into the lab to grow up two colonies of each part for minipreps on Monday morning.
Monday: Today I miniprepped the parts that I grew the previous night.
Tuesday:I ran a gel of my miniprep results from the previous day. The gel showed faint bands. I then proceeded to spec my miniprep samples to calculate the amount of DNA in each miniprep. Using this data, I did restriction digest with part B0033 in the vector backbone and C0012 as the insert.
Wednesday:I ran a gel on my restriction digest products and then proceeded with a gel extraction to obtain the vector backbone and the insert.
Thursday:Today, I redid the spec reading for my C0012 part. The new reading gave me a lower reading than the previous reading. I then redid the restriction digest for this part and ran a gel with the digest product. The gel did not show significant bands and I therefore decided to regrow the C0012 and B0033 colonies for minipreping. I conducted an ethanol precipitation and ligation with Azeem.
Friday:Today I transformed my ligation products for overnight growth. I also minipreped new colonies and ran a gel on the miniprep.
Monday:Today, I re-speced my miniprep samples from Friday using a 1:30 dilution in order to ensure that the sample level in the cuvette is high enough for the spectrometer to give an accurate reading. Using these new readings, I redid my restriction digests and ran a gel on these restriction digests. I then gel extracted the insert and the vector backbone. Finally, I grew four colonies of my ligation transformation for a miniprep.
Tuesday: I did a miniprep on the colonies I grew from last night and ran a diagnostic gel of the ligation minipreps by cutting the entire insert out of the plasmid and running a gel of the product. I also did an ethanol precipitation of the gel extraction products from the previous day and did ligations on the product. Wednesday: