Team:Ryerson Toronto/Experiments
Experiments
The genomic extraction was carried out to purify the P. aeruginosa DNA that contains the desired set of genes. In order to have a sufficient quantity of DNA, the cells were grown overnight and the DNA was released in to PBS. Following the extraction, an agarose gel electrophoresis was carried out to determine the presence of genomic DNA. The extracted DNA was diluted. Multiple dilutions were performed in order to obtain the optimum diluted conditions for the Polymerase Chain Reaction (PCR). PCR was performed using these dilutions to amplify the genes of interest using the primers that were custom designed. These primers contain the restriction enzyme sites that would be used to cleave the gene later on during transformation. Following the PCR amplification, the amplicons were visualized in a TBE gel for the correct band size. Due to difficulties in PCR, the correct band size was not observed. However, once the correct band size is obtained, the amplicons would be purified using a PCR purification kit and will be used for downstream applications.
