Since the 16S profiles in the gut appear very diverse for intracellular and extracellular DNA [33], qPCR is expected to replicate the NGS-specific Neighborhood construction both equally quantitatively and qualitatively extra closely than flow cytometry.
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A single obstacle inside the cross-research comparability of qPCR-centered quantitative microbiome profiling may be the reliance on an exterior qPCR regular from the reference organism needed to build a normal curve. In theory, any common taxon present in the microbial Group of desire can be used because the reference organism for standard curve design.
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Below, we explain how quantitative PCR (qPCR) done in parallel to NGS library preparation offers an exact estimation of complete taxon abundances from NGS details and consequently offers an attainable solution to compositionality in high-throughput microbiome analyses. The advantages and opportunity troubles of the method will also be reviewed.
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