Effect of input gDNA template concentration on microbial community composition and PUPs using DePCR
Analyses were performed on rarefied data sets (8,000 sequences per sample), with five technical replicates for each DNA input level (1.25, 2.5, 5, 10 or 20 ng/µl). (A) Genus-level mMDS ordination of microbial community structure using a distance matrix based on Bray-Curtis similarity. No significant differences were observed between all the concentrations (Global ANOSIM: R=-0.03376, p=0.79). Ellipses represent a 95% confidence interval around the centroid. (B) Primer utilization profiles for all primer variants (RPV1 – RPV18), visualized as a heatmap. (C) A positive correlation between input gDNA (1.25, 2.5, 5, 10, 20 ng/µl) and sequence yield was observed. For all input levels, the same gDNA template was used with five technical replicates. All samples were pooled after stage A of DePCR and amplified together using Illumina P5 and P7 primers. Data were rarefied to 8,000 sequences per sample.