INSA-Lyon/22 September 2010

From 2010.igem.org





September 22nd


DNA extraction of the clone of the ligation from the 21/09 of phaA/phaB


Agarose gel electrophoresis for weight verification :

  1. 1F/24A : the clone (mise en )collection is OK
  2. Curli/22B : the clone (mise en )collection is OK
  3. 8C/14K : problem with the extraction (no trace of DNA on the gel)

Digestion and purification on agarose gel:

  1. 8C, 1F/24A : with S+P
  2. 14K with X+P

Ligation overnight of :

  1. 8C/14K : one with only 14K from gel purification and one with the 8C and 14K from a gel purification
  2. 1F/24A/14K : one ligation


Measure of fluorescence and DO of the liquid culture of PHL1273 of the 20/09. Start of 6 new cultures :

  1. 3 in a water bath at 35°C, shaking speed 100 rpm
  2. 3 in a water bath at 36°C, shaking speed 100 rpm


Gel purification on vial 3 of ompR234 PCR (DNA dilution 1/10): 20µl are deposited in a lane. 100mg agarose (in 200µl NT) are extracted with NucleoSpin ExtractII kit (Macherey-Nagel). Elution was done in 50µl, and sample was stored at -20°C.


5 liquid cultures in LB medium (5ml) supplemented with corresponding antibiotic: 14K (Amp), I0500 (Kan), 1F/24A (Tet) and 18A/24A (Cm), and PHL818 (Tet), 37°C overnight.








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