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

Year 2020, Volume: 48 Issue: 1, 41 - 48, 17.04.2020

Ceren Ozkul Kocak

https://doi.org/10.15671/hjbc.538110

Abstract

The most crucial step
in molecular microbiology studies is choosing an appropriate procedure to
obtain sufficient, high-quality DNA. Two commercially available kits (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

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