User:Eric Ma/Notebook/iGEM 2010 Lab Prep/2010/05/11

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Test of old BW27783 + PBAD-RBS-RFP-Term Cultures

  • Found an old plate of BW27783s containing PBAD-RBS-RFP-Terminator in the 4ºC walk-in room
  • Tested by streaking on an Amp-100 plate + 0.1% (w/v) arabinose. Control: No arabinose
  • No growth observed on the next day.

Test of Old Antibiotic Stocks

  • Tested Amp100, Kan50 and Chlor25 from the old batch of antibiotics in the -20ºC freezer.
  • 3 ml LB + 3 µl antibiotics (each)
  • Inoculated using DH5α competent cells.
  • Results:
Amp100 Kan50 Chlor25 LB-only (Control)
No growthNo growthNo growthGrowth
  • This is a good sign. Antibiotic batches are still working.

Test of RFP-expression constructs

  • Found old Pcon-RBS-RFP-Terminator construct from the Lagally Lab -80ºC freezer. And gave a diabolical laugh when I finally found it after digging through the two -80ºC freezers we used for a whole 30 minutes.
  • Inoculated in Amp100 +/- arabinose.
  • Control: No amp, no arabinose, expect growth; next time need a negative control.
  • Setup as follows:
Reagent Tube A Tube B Tube C
Arabinose+--
Amp100++-
  • Results:
Tube Growth? Red Coloration?
AYesNo
BYesNo
CYesNo
  • No red coloration observed when expected. Therefore, I will let the cultures grow a bit more to see if red coloration appears or not.
  • Update: Red coloration was observed in tubes C (-Ara -Amp),


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