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(a) Comparison of relative abundances and approximated complete abundances of dominant bacterial families in 114 fecal samples. The best panel displays relative abundances depending on 16S amplicon sequencing as well as lower panels exhibits the believed absolute abundances calculated by multiplying the relative abundances with complete bacterial load, i.e. qPCR-dependent estimate of copies of 16S gene for every one g of feces. (b) Correlation amongst the qPCR abundances (16S rRNA gene copies for every g feces) and the believed absolute abundances of four taxa symbolizing species, genus, family members and phylum stages. The dashed line shows the anticipated 1:1 correspondence.
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qPCR is cost-efficient and obtainable because the laboratory settings, equipment and reagents are just like All those needed for preparing the NGS libraries.
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It ought to be pointed out that relative and complete abundances depending on 16S rRNA gene copies undoubtedly are a proxy for microbial density rather then exact numeration of cells, due to the fact 16S rRNA gene duplicate numbers vary among the germs. It truly is, nevertheless, feasible to computationally correct for 16S rRNA gene copy figures write-up hoc
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The same DNA extract serves because the starting up substance the two for qPCR and NGS, generating qPCR finished in 96- or 384-structure very easy to implement during the workflow for top-throughput Assessment of up to thousands of microbiome samples.
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