Virginia Commonwealth/14 July 2009
From 2009.igem.org
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Tuesday 14 July 2009
Results
Maria and Afton
- (+) Control: NEB 10 beta on LB
- lawn growth
- (+) Control: NEB 10 beta on LB shocked
- lawn growth
- (-) Control: NEB 10 beta on Cm
- no growth
- NEB 10 beta with BioBrick part (J23106, J06702, pSB4C5)
- Approximately 30 small colonies
- All of the promoters showed up on the gel electrophoresis in a mostly straight line.
Trentay 13:44, 14 July 2009 (UTC)
Kevin and Adam
- Miniprep results:
Bussingkm 15:21, 14 July 2009 (UTC)
Craig and Clay
- The ampicillin plates left overnight again displayed no growth. The electrotransformed parts will be re-plated onto ampicillin-selective plates as well as non-selective plates.
Tasks
Maria and Afton
- Digest promoters: 101, 102, 103, 104
- Possible Ligation of promoters with parts J06702, pSB4C5
Trentay 15:13, 14 July 2009 (UTC)
Kevin and Adam
- construct biobrick device (pSB4C5, BBa_E0240, BBa_J23100), and (pSB4C5, BBa_E0240, BBa_J23106)
Bussingkm 15:20, 14 July 2009 (UTC)
Craig and Clay
- The cells grown on the non-selective media will be streaked onto plates containing kanamycin and chloramphenicol to rule out possible mislabeling of resistance.
Wetlab
Maria and Afton
- Spectophotometry of promoters: 101, 102, 103, 104, 105, 107
- Digestion/Ligation of promoters
- Electrophoresis of promoter digestions: 101, 102, 103, 104, 105, 107, (100)
- Electrotransformation of ligated parts
- two Cm plate had an unidentified growth in the refrigerator.
- however, negative controls from the same plate stock show antibiotic is working
- Picked colonies for overnight culture of J23106 w/ J06702 reporter
- Made 3 overnight 5mL cultures of NEB 10 beta cell stock to prepare electrocompetent cells
Trentay 21:01, 14 July 2009 (UTC)
Kevin and Adam
- pSB4C5, BBa_E0240, and BBa_J06702 were frozen in box C-11 of the glycerol stocks
Bussingkm 15:22, 14 July 2009 (UTC)
Craig and Clay
- The cells grown on the non-selective plates were streaked onto kanamycin and chloramphenicol plates.
- Plates containing tetracycline need to be prepared to rule out this resistance.