IGEM:MIT/2006/Notebook/2006-8-2: Difference between revisions
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#Potentially LC ATF1.B0015 variants | |||
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Revision as of 14:17, 2 August 2006
to do
- LC B0030
- LC R0040
- Potentially LC ATF1.B0015 variants
- analyze 42 sequence results using prepared contigs in VectorNTI -done. unfortunately our analysis shows that nothing seems to have inserted. Perhaps digests are not being run long enough? also, pchBA mutagenesis was unsuccessful.
- start isoamyl alcohol generating device work
- PCR BAT2 and THI3 again to get higher DNA concentrations for digesting/ligating on 8/3 -done
- (3 pm) meeting with advisors -done
- run gel of PCR products/PCR cleanup and nanodrop BAT2 and THI3
- colony PCRs -done
- make liquid cultures from each potentially useful transformant plate and glycerol stocks for assembly on 8/3
meeting topics
- troubleshoot pchBA primers
- troubleshoot pchBA mutagenesis
- troubleshoot digests/failed assemblies
- timeline of assembly work (best case scenario)
liquid cultures from potentially useful plates
- 7/31 plates: 4, 5, 6, 7, 8
- 4/5/6/7 = maybe ATF1mut-Term (colony PCR to check first)
- 8 = maybe ROO40-E0840
- 8/1 plates: 7, 8, 9
- 7/8/9 = maybe ATF1mut-Term (colony PCR to check first)
liquid cultures from glycerols for assembly 8/3
primers
- design/order THI3 mutagenesis primers
- order new (add 2 more annealing bases) pchA reverse primer
team meeting: assembly plan
- current status of parts:
- WGGD: BSMT+Terminator
- SAGD: still trying to PCR pchBA
- BSGD: (?) ATF1mut+Term (?)
- IAGD: PCRed both coding regions
- osmY: pcr product