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

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#9 transformations (@2:30 in afternoon)
#9 transformations (@2:30 in afternoon)


==repeat failed digestions (and then gel extract)==
==repeat failed/funky digestions (and then gel extract)==
#R0040: SP
#R0040: SP
#RBS<sub>30A</sub>-ATF1<sub>mut</sub>: ES
#RBS<sub>30A</sub>-ATF1<sub>mut</sub>: ES

Revision as of 16:18, 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/funky digestions (and run gel)

to do (pm)

  1. gel extract digests products
  2. 8 ligations using 7/31 digested gel extracts
  3. 9 transformations (@2:30 in afternoon)

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

gel extraction pieces from 7/31

  1. RBS30A-CDS-Term: XP
  2. RBS30B-CDS-Term: XP
  3. RBS30C-CDS-Term: XP
  4. RBS32C-ATF1mut: ES
  5. RBS32A-ATF1mut: ES
  6. RBS30A-ATF1mut: ES
  7. RBS30B-ATF1mut: ES
  8. E0840: XP

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

  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

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