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

Biotechnology Reports

关键词: Droplet-digital PCR;    Quantitative real-time PCR;    Methanogen community;    Anaerobic digester;    mcrA;   

DOI  :  10.1016/j.btre.2014.06.010

来源: DOAJ

【 摘 要 】

Two different quantitative PCR platforms, droplet digital PCR (dd-PCR) and quantitative real-time PCR (qPCR), were compared in a mcrA-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−1). The mcrA-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 (R2 = 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.

【 授权许可】

Unknown