mcrA -Targeted Real-Time Quantitative PCR Method To Examine Methanogen Communities
نویسندگان
چکیده
منابع مشابه
mcrA-targeted real-time quantitative PCR method to examine methanogen communities.
Methanogens are of great importance in carbon cycling and alternative energy production, but quantitation with culture-based methods is time-consuming and biased against methanogen groups that are difficult to cultivate in a laboratory. For these reasons, methanogens are typically studied through culture-independent molecular techniques. We developed a SYBR green I quantitative PCR (qPCR) assay...
متن کاملComparison 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 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 c...
متن کاملQuantitative Real-Time PCR
Unlike classical end-point analysis PCR, real-time PCR provides the data required for quantification of the target nucleic acid. The results can be expressed in absolute terms by reference to external quantified standards or in relative terms compared to another target sequence present within the sample. Absolute quantification requires that the efficiency of the amplification reaction is the s...
متن کاملReal time quantitative PCR.
We have developed a novel "real time" quantitative PCR method. The method measures PCR product accumulation through a dual-labeled fluorogenic probe (i.e., TaqMan Probe). This method provides very accurate and reproducible quantitation of gene copies. Unlike other quantitative PCR methods, real-time PCR does not require post-PCR sample handling, preventing potential PCR product carry-over conta...
متن کاملReal-time quantitative PCR.
Those who have worked in the field of quantitative polymerase chain reaction (PCR) since the early 1990s have accepted many of the tedious aspects of the early assays as routine. Difficulties associated with early quantitative PCR techniques included: (i) ensuring that the PCR was within the linear range of amplification (that portion where the PCR signal is directly proportional to the input c...
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ژورنال
عنوان ژورنال: Applied and Environmental Microbiology
سال: 2009
ISSN: 0099-2240,1098-5336
DOI: 10.1128/aem.02858-08