COMPARISON OF BACTERIAL DNA EXTRACTION METHODS FROM STOOL SAMPLES FOR QUANTITATIVE REAL-TIME PCR ANALYSIS

Authors : Ceren Ozkul KOCAK

Pages : 41-48

Doi:10.15671/hjbc.538110

View : 17 | Download : 16

Publication Date : 2020-04-17

Article Type : Research Paper

Abstract :The most crucial step in molecular microbiology studies is choosing an appropriate procedure to obtain sufficient, high-quality DNA. Two commercially available kits insert ignore into journalissuearticles values(QIAamp DNA Stool Mini Kit and PSP Spin Stool DNA Plus Kit); and a modified in-house phenol-chloroform extraction method was compared for their efficiency in isolation of certain bacteria from human stool samples for Real-Time qPCR. DNA yield was significantly higher in Invitrogen when compared to phenol-chloroform method. Spiking experiments and qPCR results revealed that efficiency was in phenol-chloroform extraction was lower when compared to other two commercial kits. Commercial kits have better results in terms of DNA recovery and purity, and have more accurate results for qPCR experiments.
Keywords : qPCR, DNA extraction, stool, phenol chloroform