Endpoint Recombinase Polymerase Amplification (RPA) Assay for Enumeration of Thiocyanate-degrading Bacteria

Mamoru Oshiki, Toshikazu Fukushima, Shuichi Kawano, Junichi Nakagawa

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

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.

Original languageEnglish
Article numberME21073
JournalMicrobes and environments
Volume37
Issue number1
DOIs
Publication statusPublished - 2022
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Ecology, Evolution, Behavior and Systematics
  • Soil Science
  • Plant Science

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