IGEM:MIT/2006/Notebook/2006-8-2: Difference between revisions
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==to do== | ==to do== | ||
#LC B0030 | |||
#LC R0040 | |||
#LC B0015 | |||
#LC ATF1.B0015 variants | |||
#LC R0040.E0840 | |||
<br><br> | |||
#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. | #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 | #start isoamyl alcohol generating device work | ||
#*PCR BAT2 and THI3 again to get higher DNA concentrations for digesting/ligating on 8/3 | #*PCR BAT2 and THI3 again to get higher DNA concentrations for digesting/ligating on 8/3 -'''done''' | ||
#(3 pm) meeting with advisors | #(3 pm) meeting with advisors -'''done''' | ||
#run gel of PCR products/PCR cleanup and nanodrop BAT2 and THI3 | #run gel of PCR products/PCR cleanup and nanodrop BAT2 and THI3 | ||
#make liquid cultures | #colony PCRs -'''done''' | ||
#make liquid cultures from each potentially useful transformant plate and glycerol stocks for assembly on 8/3 | |||
==meeting topics== | ==meeting topics== | ||
#troubleshoot pchBA primers | #troubleshoot pchBA primers | ||
#troubleshoot pchBA mutagenesis | #troubleshoot pchBA mutagenesis | ||
#troubleshoot digests/failed assemblies | |||
#timeline of assembly work (best case scenario) | #timeline of assembly work (best case scenario) | ||
==liquid cultures from potentially useful plates== | ==liquid cultures from potentially useful plates== | ||
#7/31 plates: 4, 5, 6, 7, 8 | #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 | #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 |
Latest revision as of 15:03, 2 August 2006
to do
- LC B0030
- LC R0040
- LC B0015
- LC ATF1.B0015 variants
- LC R0040.E0840
- 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