title: Development of Methanoculleus-specific real-time quantitative PCR assay for assessing methanogen communities in anaerobic digestion.
abstract: OBJECTIVE
  To develop a Methanoculleus-specific real-time quantitative PCR (RT-qPCR) assay with high coverage and specificity for the analysis of methanogenic populations in anaerobic digestion.
  
  
  RESULTS
  A Methanoculleus-specific primer/probe set for RT-qPCR was designed in this study based on all Methanoculleus 16S rRNA gene sequences in Ribosomal Database Project (RDP) according to TaqMan chemistry. The newly designed primer/probe set was shown to have high coverage and specificity by both in silico and experimental analyses. Amplification efficiency of the Methanoculleus-specific primer/probe set was determined to be ideal for RT-qPCR applications. Subsequent field testing on anaerobic digesters showed that results from RT-qPCR were consistent with those from clone library analysis, validating the accuracy of the RT-qPCR assay.
  
  
  CONCLUSIONS
  The Methanoculleus-specific RT-qPCR assay designed in this study can serve as a rapid and effective tool for the quantification of Methanoculleus populations in anaerobic digestion.
  
  
  CONCLUSIONS
  Methanoculleus populations represent important members of archaeal communities in methanogenic processes, necessitating the need to develop effective tools to monitor Methanoculleus population abundance. The RT-qPCR developed in this study provides an essential tool for the quantification of Methanoculleus populations in anaerobic digestion and for the understanding of the functions of these methanogens in anaerobic biotransformation.
authors: Department of Civil and Environmental Engineering, The University of Tennessee, Knoxville, TN, USA.
authors: S
authors: S
authors: Chen
authors: Z
authors: Z
authors: Zhu
authors: J
authors: J
authors: Park
authors: Z
authors: Z
authors: Zhang
authors: Q
authors: Q
authors: He
mesh_terms: D019149:Bioreactors / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics*; D019638:Methanomicrobiaceae / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D012336:RNA, Ribosomal, 16S
keywords: Methanoculleus; anaerobic digestion; methanogen; quantitative PCR; real-time PCR