840:153g:Projects/project18/2011/09/22

Cloning of catA gene of Acinetobacter calcoaceticus strain ADP1 to observe its expression in E.coli.

Thursday September 22, 2011. Transformation of the Plasmid DNA of our promoter of choice BBa_K118011 and PsB1A7 DNA given by Axel was done. The required materials for Transformation were Competent cells, Plasmid DNA, SOC media, LB media plates. Firstly, Competent cells and Plasmid DNA was thawed on ice. Then, five tubes were taken. One of the tube had control which is competent cells and other were DNA samples of BBa_K118011 and PsB1A7 of different concentration. To these samples 50ul of competent cells were added and thawed on ice for 20 mins. In the meanwhile water bath was set to 42 C. After thawing on ice, samples were given heat shock for 50 secs at 42C then kept on ice for 2 mins. 1 ml of SOC media was added to all the tubes and kept in incubator shaker for an hour at 37C to increase the transformation efficiency of the Plasmid. After an hour the tubes were taken from the incubator shaker, 5 were plated with 200ul of sample in ampicillin plates and as control for our transformation two other plates were taken and 50ul of competent cells were added one with ampicillin and one without ampicillin to check the growth of competent cells. After transformation was done, plates were kept in incubator at 37c, overnight for bacteria to grow. By Sanjana Gurrala.