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

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) solutio...

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Detalles Bibliográficos
Autores principales: Oshiki, Mamoru, Fukushima, Toshikazu, Kawano, Shuichi, Nakagawa, Junichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Japanese Society of Microbial Ecology / Japanese Society of Soil Microbiology / Taiwan Society of Microbial Ecology / Japanese Society of Plant Microbe Interactions / Japanese Society for Extremophiles 2022
Materias:
Short Communication
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8958297/
https://www.ncbi.nlm.nih.gov/pubmed/35264493
http://dx.doi.org/10.1264/jsme2.ME21073
Descripción
Sumario: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 2.5×10(3) and 1.5×10(6) copies μL(–1), showing a linear relationship (R(2)=0.83) with those measured using a conventional quantitative PCR assay.

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