June 24th, 2010
I purified the PCR product I obtained yesterday containing LuxAB with High Pure PCR product Purification Kit by Roche.
Then I made an agarose gel to get sure that I didn't lose the PCR product during the purification.
The gel looked hazy but the PCR product was there.
Then I made the following restrictions:
LuxAB obtained from PCR prodcut with XbaI and PstI
Plasmid 17 with RFP with XbaI and PstI
Plasmido with constitutive promoter (J23106) SpeI and PstI
I also searched for protocols to use decanal for luminescence tests:
50μL bacterial culture they add:
aliquots: 20μL decanal (a range of different concentrations from 2.5 to 17.4mM in 1:1 distilled water - methanol)
aliquot: 500μL (13.2μM decanal, 1% ethanol)
Then I inactivated restriction enzymes by a heat-shock (80°C during 20min) and I made an agarose gel 0.8% preparing ligation.