IGEM:MIT/2006/Notebook/2006-8-2

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(to do)
Current revision (18:03, 2 August 2006) (view source)
(to do)
 
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#LC B0030
#LC B0030
#LC R0040
#LC R0040
 +
#LC B0015
#LC ATF1.B0015 variants
#LC ATF1.B0015 variants
#LC R0040.E0840
#LC R0040.E0840

Current revision

Contents

to do

  1. LC B0030
  2. LC R0040
  3. LC B0015
  4. LC ATF1.B0015 variants
  5. LC R0040.E0840



  1. 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.
  2. start isoamyl alcohol generating device work
    • PCR BAT2 and THI3 again to get higher DNA concentrations for digesting/ligating on 8/3 -done
  3. (3 pm) meeting with advisors -done
  4. run gel of PCR products/PCR cleanup and nanodrop BAT2 and THI3
  5. colony PCRs -done
  6. make liquid cultures from each potentially useful transformant plate and glycerol stocks for assembly on 8/3

meeting topics

  1. troubleshoot pchBA primers
  2. troubleshoot pchBA mutagenesis
  3. troubleshoot digests/failed assemblies
  4. timeline of assembly work (best case scenario)

liquid cultures from potentially useful plates

  1. 7/31 plates: 4, 5, 6, 7, 8
    • 4/5/6/7 = maybe ATF1mut-Term (colony PCR to check first)
    • 8 = maybe ROO40-E0840
  2. 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

  1. design/order THI3 mutagenesis primers
  2. order new (add 2 more annealing bases) pchA reverse primer

team meeting: assembly plan

  1. current status of parts:
    • WGGD: BSMT+Terminator
    • SAGD: still trying to PCR pchBA
    • BSGD: (?) ATF1mut+Term (?)
    • IAGD: PCRed both coding regions
    • osmY: pcr product
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