IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/06/09

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Biofilm Track

Eric F.

  • Recieved S. Aureus from the Davies lab on a streaked plate. Strain is currently unknown, will know soon (through Rafael)
  • Used the Day Culture protocol from common protocols
  • Picked two colonies into two 14mL test tubes each containing 5mL LB broth
  • Made one control containing just 5mL broth
  • Incubated all 3 tubes at 37 degrees C for 9 hours
  • Control remain clear, others were very cloudy
  • Now being stored on the bench in a rack

QS Track

Jason and Eric F.

Competent Cell Transformation

  • Tested the competent cells (DH5α and Db3.1 - made by Philip and Kevin) made yesterday using plasmid from last year labeled ASEM24(B) with Ampicilin resistance
    • Tested one from each centrifuge tube, plated labeled B1, B2, B3, α1 ,α2
  • Used the transformation protocol from common protocols
    • In step 1, the competent cells were allowed to thaw for 1 hour on ice, and were occasionally hand warmed
    • During step 2, noticed the α2 aliquot had too much volume, so we put it back in the -80 degree C and got a different one
      • Note: This aliquot was hand heated more than the others and was only thawed on ice for 10 minutes
  • 5 plates of transformed cells were spread onto Ampicilin plates and are incubating overnight in the biosafety room incubator at 37 degrees C
    • Need to be removed by 1pm tomorrow
  • Note: Need to plate more Amp plates tomorrow
  • Note: There was no control, if there is no transformation, we should test the DNA with last year's competent cells

PCR Previously Transformed Cells

  • Obtained results from the PCR of the transformed cells
  • Results were inconclusive and confusing, the ladders showed no movement, only the dye moved (the dye moves independently right?)
    • Picture is in the QS Track labbook
    • Had trouble getting a clear picture

Misc.

Eric F.

  • Removed culture of BW27783 from the incubator (30 degree) at 10am
  • Control remained clear, tube containing E. Coli was quite cloudy
  • Need to send to Tec-Monterrey, tubes are on the bench in a rack





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