IGEM:MIT/2006/Notebook/2006-9-1

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(to do)
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#*J45220-C-mut with ATF1 primers
#*J45220-C-mut with ATF1 primers
#*all other minipreps (except pE3)
#*all other minipreps (except pE3)
-
#*J45298-no mut: 3,4,6,7
+
#*J45398-no mut: 3,4,6,7
#PCR: pmsCEAB out of pE3(new) and pE3R  
#PCR: pmsCEAB out of pE3(new) and pE3R  
#*adjust annealing time, check GC content, ask advisor advice on PCRing large coding regions
#*adjust annealing time, check GC content, ask advisor advice on PCRing large coding regions

Revision as of 14:26, 1 September 2006

to do

  1. glycerol: all LCs
  2. start a smell test: add precursor to the LCs of 0-Q-119 and O-Q-199
  3. am miniprep (12): J45397 (1), J45398 (1), J45399 (3), J45299 (3), J45319 (3), pE3 (1)
    • note: I don't think that the yeast will be grown up enough to miniprep this early in the day
  4. analyze: incoming 20 sample sequencing order
    • throw out bad sequences
  5. digest: J45299 (3) and J45319 (3) with XP
    • run a gel
  6. sequence: (32 wells)
    • J45220-C-mut with ATF1 primers
    • all other minipreps (except pE3)
    • J45398-no mut: 3,4,6,7
  7. PCR: pmsCEAB out of pE3(new) and pE3R
    • adjust annealing time, check GC content, ask advisor advice on PCRing large coding regions
    • clean up??
    • run a gel
  8. graph the plate reader data (call stephen)
  9. 3-part ligate/transform into A/C backbone:
    • J45299:XP with R0011:ES
    • J45319:XP with R0011:ES
    • need to find more backbone!!
  10. pellet: yeast cells in big centrifuge, resuspend in milk, try baking
    • ask samantha for help
  11. pm miniprep: iGEM yeast
    • follow yeast miniprep protocol, vortex cells first, ask samantha for help
    • we can do XS digest/freeze of yeast miniprep on monday and ligate/transform with ATF1 when samantha is around and when we have tried out the bread machine
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