Optimization of DNA extraction for obesity study / Farhah Nadia Zulkifli

Quality and quantity of DNA is crucial for downstream analysis in nucleic acids research especially with raising interest in pharmacogenomics study. Commercially available DNA extraction kit would include protocols and procedure for the kit, however it could be optimized through adjustment of certain parameters to obtain better yield and greater quality of DNA. The aim of this study is to improve quality and quantity of DNA obtained. For that, adjustments were made on centrifugation speed and incubation time to determine their influence on DNA yield. Standard procedure for centrifuge speed was 14000 rpm for 1 minute was modified to 7000 rpm for 20 minutes and incubation time was increased twice the time of suggested protocol from Promega. It was observed that yield of samples processed with modified procedure was statistically significant increase from a mean of 35.22 ng/ml to 69.97ng/ml. Even with statistically significant data (p=0.49<0.05), a conclusive remarks could not be generated due small sample size (n=6). In order to validate the data, the sample size should be increased and several centrifugation speeds are chosen to further optimize DNA extraction.