학술논문

Endpoint Recombinase Polymerase Amplification (RPA) Assay for Enumeration of Thiocyanate-degrading Bacteria
Document Type
Journal Article
Source
Microbes and Environments. 2022, 37(1):21073
Subject
endpoint assay
gene quantification
recombination amplification reaction (RPA)
thiocyanate dehydrogenase
Language
English
ISSN
1342-6311
1347-4405
Abstract
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.