ULB/20 August 2009

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Contents

August 20, 2009

Plasmid extraction

Method

Plasmids have been extracted from our bacteria cultures.

  • ccdA forward : 2 tubes
  • promoter + RBS + RFP : 4 tubes, half of them contained red bacteria

Results

Photometry showed that there is very few extracted genetic material, except for the first ccdA tube. New cultures have then been setup for the promoter + RBS + RFP plasmid for a new extraction.

Photo missing for gel migrations

Results from the gel migration have not yet been interpreted.

No glycerol has been done.

Ligations: hfsG + hfsH + terminator

hfsG and hfsH are located on plasmids carrying Amp resistance from Mr. Gene. In order to be able to select plasmids with antibiotics, we will first link hfsH with our terminator because the plasmid carrying the terminator is resistant to Kanamycin (and Amp).

hfsH was digested with EcoRI and SpeI. terminator was digested with EcoRI and XbaI. The rest of the digestion follows the standard protocol. The result was dialysed and ligated.

Solid culture boxes have been created with Kanamycin.

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