Cloning of catA gene of Acinetobacter calcoaceticus strain ADP1 to observe its expression in E.coli.
Today we had run the 1% agaraose gel electrophoresis of our genomic DNA that we obtained on 10/27/2011. The main purpose was to check the amplification of the desired gene.We have used 12 wells in the gel in which first wel was 100bp DNA ladder ,second well was empty, third well was negative control,fourth well was positive control and from well five to twelve were the DNA sample.Gel was run for 30 mins at 150 volts.There was band formed in well 4 ,which shows that positive control was amplified and as expected there is no band formation in well 3 which is negative control. There are no band formations from well 5-12 ,showing that there was no DNA amplification. This could be due to primers we used or the bacterial growth se we would be repeating the bacterial growth step. So we rehydrated the Acinetobacter Calcoaceticus strain ADP1 ,so that we can work with fresh transformed plates.We also extracted the promoter parts from iGem distribution kits of BBa_1765001 and BBa_k118011 of 2010 and 2009.We extracted the same promoter part we used earlier because it gave good result but from previous year kit as we have already used it from 2011 kit.And in the end we made LB media plates that we will autoclave on thursday and use it for promoter part transformation.