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Considering that many common bacterial primers have been designed and optimized especially for qPCR or NGS, it is actually a good idea to take into account prospective biases ensuing from primer-unique amplification effectiveness for unique taxa [35] and differential primer protection, when working with distinct primer sets for qPCR and NGS. In this particular study, we chose a broadly-employed common bacterial primer established optimized for qPCR [29] that similarly targets the V3-V4 hypervariable regions since the primer set utilized for Illumina MiSeq (S1A Table).
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The sequencing reads assigned to different taxa in Each and every sample were being divided by the total number of reads for the sample to obtain relative abundances of your taxa in each sample. The relative abundances obtained depending on the sequencing reads have been translated into whole abundances by multiplying the relative abundance of each and every taxon by the entire bacterial abundance in the sample.
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