IGEM06 DNA Distribution Issues
From 2006.igem.org
| Line 67: | Line 67: | ||
|DH5a, Rosetta | |DH5a, Rosetta | ||
|iGEM2006 DNA-1 | |iGEM2006 DNA-1 | ||
| - | | We have had persistent issues with our cells being unable to express this protein (AHL-Lactonase). Upon sequencing our constructs it appears that the DNA we received from the registry is an empty vector with no aiiA gene present. Our sequencing results can be seen in this report. | + | | We have had persistent issues with our cells being unable to express this protein (AHL-Lactonase). Upon sequencing our final constructs it appears that the DNA we received from the registry is an empty vector with no aiiA gene present. Our sequencing results can be seen in [this report.] This is a tentative announcement as we are still waiting for sequencing results from the wells. |
It is very important that these results are checked and verified as there are 15 available (non-working) parts which rely on this gene and 74+ parts which have been designed/built but not submitted to the registry. | It is very important that these results are checked and verified as there are 15 available (non-working) parts which rely on this gene and 74+ parts which have been designed/built but not submitted to the registry. | ||
Revision as of 16:00, 12 September 2006
This page is to keep a record of any troubles that you have when trying to successfully transform parts from either of the two iGEM 2006 DNA Distribution plates (See Biobrick Delivery page for delivery details)
Please include:
- Biobrick part number
- Plasmid
- Cell strain used
- Plate (iGEM 2006 DNA-1 or DNA-2) and well number
| Team | Part number | Plasmid | Cell strain | Distribution plate | Comments |
|---|---|---|---|---|---|
| Example | BBa_xxxx | pSB1A2 | TOP10 | iGEM06 DNA-1 | we tried 10 times!! |
| PennState | BBa_R0051 | pSB1A2, pSB2K3 | DH5a | iGEM06/05 DNA-1 | neither amp plate seems to work |
| Davidson | BBa_B0015 | pSB1AK3 | JM109 | iGEM06 DNA-1 | no insert at all; had to use stocks from last year; pSB1A2 |
| Berkeley | BBa_P1000 (CmR) | pSB1AC3 | TG1 | iGEM06 DNA-2 | Austin says it should be a pSB2K* plasmid; no CmR colonies generated upon transformation |
| Berkeley | BBa_P1001 (tet) | pSB1AT3 | TG1 | iGEM06 DNA-2 | Austin says it should be a pSB2K* plasmid; no tetR colonies generated upon transformation |
| Ljubljana, Slovenia | BBa_P1010 (tet) | pSB1AT3 | DB3.1 | iGEM06 DNA-2 (23P) | Plasmid pSB1AT3 should have tet resistance, but instead it has Kan resistance (so it should be labeled pSB1AK3). |
| Cambridge | BBa_J04430 | pSB1A2 | DH5-a; XL-1 Blue; MG-1655; MC1000 | iGEM06 DNA-2 (1L) | On transforming, this biobrick persistently results in segregated colonies, some show the expected green fluorescence but others show surprisingly red fluorescence. IPTG does not have an effect. This has been our experience |
| Imperial | BBa_C0060
and all other bricks containing aiiA | pSB1A2 | DH5a, Rosetta | iGEM2006 DNA-1 | We have had persistent issues with our cells being unable to express this protein (AHL-Lactonase). Upon sequencing our final constructs it appears that the DNA we received from the registry is an empty vector with no aiiA gene present. Our sequencing results can be seen in [this report.] This is a tentative announcement as we are still waiting for sequencing results from the wells.
It is very important that these results are checked and verified as there are 15 available (non-working) parts which rely on this gene and 74+ parts which have been designed/built but not submitted to the registry. List of potentially affected parts which have been distributed
|
If we see the same Biobrick parts giving multiple teams trouble it will give us a better sense of what we need to fix.
Note: Please take a look and see if your team has been able to successfully transform any of the parts on this list. You might want to contact the team that was having trouble and let them know that you were able to successfully transform that particular part. All of the distribution plates were done in the same method at the same time so they should be the same, but that isn't always the case.
