Abstract:
Objectives: Extraction of DNA and RNA is the first step in genomics and transcriptomics studies. Phenolchloroform method for DNA extraction has been the widely used method. However, this method is relatively expensive and time-consuming. The objective of the present study was to validate a cost and timeeffective protocol that will reduce the burden of molecular biology-based research and make a difference in laboratories with limited resources. Methods: A comparative study was conducted at Syed Qamer Alam Research Laboratory, Shifa College of Medicine; from February, 2021 to August, 2021. TRIzolTM method was used to extract RNA from blood samples of coronary artery disease patients and remnant was used to extract DNA. The quantity, purity and integrity of the extracted DNA by both methods (TRIzol and phenol-chloroform) was examined. PCR product amplification was performed with thrombomodulin (THBD) gene to validate the characteristic of the extracted DNA and its efficiency for downstream experiments. Results: The DNA yield in the TRIzolTM method was three-fold higher than phenol chloroform method. Both methods showed intact genomic DNA on the agarose gel, and extracted DNA was efficient for PCR amplification. Conclusion: The TRIzolTM method for RNA and DNA co-extraction is fast, simple and economical technique. So, it can be adopted for routine molecular biology analyses in limited resources setup.
Page(s):
1754-1759
DOI:
DOI not available
Published:
Journal: Pakistan Journal of Medical Sciences, Volume: 38, Issue: 7, Year: 2022
Keywords:
Coronary artery disease
,
DNA extraction
,
RNA extraction
,
Costeffective
,
Coextraction