Isolation of nucleic acids from feces using a silica-based sorbent for the identification of new probiotic agents
Abstract
One of the main stages of molecular genetic research is the extraction of nucleic acids. The sorption-based DNA/RNA isolation technique has proven to be a successful and popular technology. The isolation of nucleic acids from complex substrates such as feces can be a problem. Feces are known to contain novel probiotic agents such as Faecalibacterium prausnitzii and Akkermansia muciniphila, which have a wide range of biological activities. The work is devoted to evaluating the effectiveness of DNA and RNA extraction from feces using silica (silicon dioxide) as a sorbent for nucleic acids, with the addition of Triton X-100, Tween 20, Tween 80 and Pluronic P123 as a detergent in the lysing solution in concentrations of 1, 3, 5 and 10%. Electrophoresis in agarose gel showed that an increase in the concentration of detergent provoked an increase in the intensity of the glow bands characteristic of both DNA and RNA. The highest RNA concentrations were typical for 10% Tween 80 and 5% Triton X-100. At the same time, with an increase in the concentration of detergent, a greater RNA yield was observed. The maximum values of DNA concentration were observed using Tween 20 as detergent. The data are consistent with the results obtained based on Real-Time PCR. It has been shown that the use of Tween 20 detergent is most optimal for DNA isolation from fecal bacteria Faecalibacterium prausnitzii and Akkermansia muciniphila.
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