Because the 16S profiles through the gut appear quite unique for intracellular and extracellular DNA [33], qPCR is anticipated to mirror the NGS-focused Neighborhood framework both quantitatively and qualitatively extra carefully than flow cytometry.
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Importantly, qPCR-based quantitative microbiome profiling enjoys the following conceptual and functional Added benefits above other approaches:
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The use of relative abundance info from next technology sequencing (NGS) can result in misinterpretations of microbial more info Neighborhood structures, as the increase of one taxon contributes to the concurrent lower of the other(s) in compositional facts. While distinct DNA- and cell-based procedures together with statistical strategies happen to be designed to beat the compositionality dilemma, along with the Organic relevance of complete bacterial abundances has been demonstrated, the human microbiome exploration has not still adopted these solutions, probably resulting from feasibility difficulties.
Below, we describe how quantitative PCR (qPCR) accomplished in parallel to NGS library planning provides an accurate estimation of absolute taxon abundances from NGS facts and consequently offers an attainable Resolution to compositionality in higher-throughput microbiome analyses. The benefits and opportunity problems of the method are also mentioned.
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