Abstract
Development of an Optimized Protocol for the Extraction of DNA from Human Blood for the Isolation and Expression of the Human Gene CtsK, in Bacterial Expression System, Escherichia Coli
Madubashetha, H., Dissanayake, Y., Piyasiri, L., Wickramasinghe, S., De Silva, N., Warnakula, L. & Cooray, R.
11th October 2019
Life Sciences, 6th PGIS Research Congress 2019 (RESCON 2019), University of Peradeniya, Peradeniya, Sri Lanka.
doi:
Cathepsin K encoded by human gene CtsK, is a major enzyme involved in bone remodeling. The study was built to investigate the feasibility of in vitro expression of CtsK gene in bacterial expression system. The original protocol for DNA extraction from human blood given in FlexiGene® DNA extraction kit by QIAGEN® was optimized. As modifications to the original protocol, volume and number of isopropanol and ethanol purification steps, air-drying time were increased which led to improved A260/A280 and A260/230 ratios and increased DNA concentration.