IGEM:MIT/2006/Notebook/2006-8-1: Difference between revisions

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#test osmY+E0840(A,B,C) part with plate reader
#test osmY+E0840(A,B,C) part with plate reader
#miniprep/nanodrop 7 overnight liquid cultures
#miniprep/nanodrop 7 overnight liquid cultures
#repeat failed/funky digestions (and run gel --NO GEL EXTRACTION)
#repeat failed 7/31 digestions


==to do (pm)==
==to do (pm)==

Revision as of 16:50, 31 July 2006

to do (am)

  1. ask Isadora to help with fax machine
  2. order new pchA reverse primer (ask Tom for help?)
  3. run a gel of BAT2 and THI3 pcr products
  4. test osmY+E0840(A,B,C) part with plate reader
  5. miniprep/nanodrop 7 overnight liquid cultures
  6. repeat failed 7/31 digestions

to do (pm)

  1. nanodrop and run a gel of digest products (NO GEL EXTRACTION)
  2. do ligations using new 8/1 digest products
  3. transform

repeat failed/funky digestions (and then gel extract)

  1. R0040: SP
  2. RBS30A-ATF1mut: ES
  3. RBS30B-ATF1mut: ES
  4. RBS30C-ATF1mut: ES
  5. RBS32A-ATF1mut: ES
  6. RBS32B-ATF1mut: ES
  7. RBS32C-ATF1mut: ES


other cut pieces we have ready to use

  1. B0015: EX
  2. B0030: SP
  3. B0032: SP
  4. R0040:SP (didn't show with 5μL loaded on wide lane gel, but could still be there)

ligations (with 7/31 gel extract pieces 1-8 above)

  1. R0040+1->Prom-RBS30A-CDS-Term
  2. R0040+2->Prom-RBS30B-CDS-Term
  3. R0040+3->Prom-RBS30C-CDS-Term
  4. 4+Term->RBS32C-ATF1mut-Term
  5. 5+Term->RBS32A-ATF1mut-Term
  6. 6+Term->RBS30A-ATF1mut-Term
  7. 7+Term->RBS30B-ATF1mut-Term
  8. R0040+8(E0840)->CP-RBS-GFP-Term

transformations (@ 2:30 pm)

  1. prepare 9 plates, review the resistance of backbones
  2. use fresh Top10 cells, pUC19 control, 200 μL
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