TY - JOUR
T1 - Endpoint Recombinase Polymerase Amplification (RPA) Assay for Enumeration of Thiocyanate-degrading Bacteria
AU - Oshiki, Mamoru
AU - Fukushima, Toshikazu
AU - Kawano, Shuichi
AU - Nakagawa, Junichi
N1 - Publisher Copyright:
© 2022, Japanese Society of Microbial Ecology. All rights reserved.
PY - 2022
Y1 - 2022
N2 - An endpoint recombination amplification reaction (RPA) assay for assessing the abundance of the gene encoding thiocyanate dehydrogenase (TcDH) in Thiohalobacter has been developed. The RPA reaction was performed at 37°C for 30 min, terminated by the addition of sodium dodecyl sulfate (SDS) solution, and the DNA concentration of the RPA product was fluorometrically measured. The abundance of TcDH in 22 activated sludge samples and 7 thiocyanate-degrading enrichment cultures ranged between 2.5×103 and 1.5×106 copies μL–1, showing a linear relationship (R2=0.83) with those measured using a conventional quantitative PCR assay.
AB - An endpoint recombination amplification reaction (RPA) assay for assessing the abundance of the gene encoding thiocyanate dehydrogenase (TcDH) in Thiohalobacter has been developed. The RPA reaction was performed at 37°C for 30 min, terminated by the addition of sodium dodecyl sulfate (SDS) solution, and the DNA concentration of the RPA product was fluorometrically measured. The abundance of TcDH in 22 activated sludge samples and 7 thiocyanate-degrading enrichment cultures ranged between 2.5×103 and 1.5×106 copies μL–1, showing a linear relationship (R2=0.83) with those measured using a conventional quantitative PCR assay.
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U2 - 10.1264/jsme2.ME21073
DO - 10.1264/jsme2.ME21073
M3 - Article
C2 - 35264493
AN - SCOPUS:85126389314
SN - 1342-6311
VL - 37
JO - Microbes and environments
JF - Microbes and environments
IS - 1
M1 - ME21073
ER -