Comparison of droplet digital PCR and quantitative real-time PCR in mcrA -based methanogen community analysis

Two different quantitative PCR platforms, droplet digital PCR (dd-PCR) and quantitative real-time PCR (qPCR), were compared in a -based methanogen community assay that quantifies ten methanogen sub-groups. Both technologies exhibited similar PCR efficiencies over at least four orders of magnitude an...

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Veröffentlicht in:Biotechnology reports (Amsterdam, Netherlands) Netherlands), 2014-12, Vol.4 (C), p.1-4
Hauptverfasser: Kim, Tae Gwan, Jeong, So-Yeon, Cho, Kyung-Suk
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Sprache:eng
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Zusammenfassung:Two different quantitative PCR platforms, droplet digital PCR (dd-PCR) and quantitative real-time PCR (qPCR), were compared in a -based methanogen community assay that quantifies ten methanogen sub-groups. Both technologies exhibited similar PCR efficiencies over at least four orders of magnitude and the same lower limits of detection (8 copies μL-DNA extract ). The -based methanogen communities in three full-scale anaerobic digesters were examined using the two technologies. dd-PCR detected seven groups from the digesters, while qPCR did five groups, indicating that dd-PCR is more sensitive for DNA quantification. Linear regression showed quantitative agreements between both of the technologies (  = 0.59-0.98) in the five groups that were concurrently detected. Principal component analysis from the two datasets consistently indicated a substantial difference in the community composition among the digesters and revealed similar levels of differentiation among the communities. The combined results suggest that dd-PCR is more promising for examining methanogenic archaeal communities in biotechnological processes.
ISSN:2215-017X
2215-017X
DOI:10.1016/j.btre.2014.06.010