Rapid enumeration of sulphate-reducing bacteria from aquatic environments using real-time PCR
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- Kondo Ryuji
- Department of Marine Bioscience, Fukui Prefectural University
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- Shigematsu Kotaro
- Graduate School of Bioscience and Biotechnology, Fukui Prefectural University
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- Butani Junki
- Graduate School of Bioscience and Biotechnology, Fukui Prefectural University
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We describe a rapid and simple enumeration method for sulphate-reducing bacteria (SRB) from aquatic environments using a quantitative real-time PCR (qPCR) in combination with a rapid DNA isolation method. Enumeration of SRB in the sediment and water samples was performed by quantifying the copy number of the dsrA gene coding for the α-subunit of the dissimilatory sulphite reductase using real-time PCR with the SYBR Green I assay. Using dsrA-specific primers, we demonstrated that quantification of SRB in known numbers of SRB assemblages can be achieved. We compared DNA isolation methods using commercial DNA extraction kits and a published technique. We found two commercial kits were of advantage for extraction of DNA from water or sediment samples, where a large number of samples could be processed at the same time. We showed this newly developed qPCR technique targeting dsrA is rapid, simple and reproducible for the quantification of SRB numbers in situ and is superior to the use of culture-dependent techniques.
収録刊行物
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- Plankton and Benthos Research
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Plankton and Benthos Research 3 (3), 180-183, 2008
日本プランクトン学会、日本ベントス学会
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詳細情報 詳細情報について
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- CRID
- 1390282680242084096
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- NII論文ID
- 10021907601
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- NII書誌ID
- AA12130745
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- ISSN
- 1882627X
- 18808247
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- NDL書誌ID
- 9638127
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- 抄録ライセンスフラグ
- 使用不可