Use of artificial DNA with multiple probe sites as reference DNA templates for quantitative real-time PCR to examine methanogen communities

Tae Gwan Kim, Taewoo Yi, Kyung Suk Cho

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Isolation of reference DNA templates for quantitative real-time PCR assays is an expensive, labor-intensive and time-consuming process if they are not readily available. Two artificial DNA templates with multiple probe sites were designed for quantifying methanogens and their 10 subgroups, based on the methyl coenzyme M reductase gene (mcrA). Their standards were comparable to each other. PCR amplification efficiencies (cycle vs. cumulative fluorescence) of the artificial DNAs were also comparable to those of the observed methanogen groups from anaerobic digesters. The artificial templates can be alternatives to the actual references.

Original languageEnglish
Pages (from-to)417-421
Number of pages5
JournalJournal of Environmental Science and Health - Part A Toxic/Hazardous Substances and Environmental Engineering
Volume48
Issue number4
DOIs
StatePublished - 1 Mar 2013

Keywords

  • Quantitative PCR
  • artificial DNA
  • mcrA
  • methanogens

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