biostudies-literatureUnknown6(10)Manzari CMinistero dell’Istruzione, dell’Università e della RicercaThe quantification of the total microbial content in metagenomic samples is critical for investigating the interplay between the microbiome and its host, as well as for assessing the accuracy and precision of the relative microbial composition which can be strongly biased in low microbial biomass samples. In the present study, we demonstrate that digital droplet PCR (ddPCR) can provide accurate quantification of the total copy number of the 16S rRNA gene, the gene usually exploited for assessing total bacterial abundance in metagenomic DNA samples. Notably, using DNA templates with different integrity levels, as measured by the DNA integrity number (DIN), we demonstrated that 16S rRNA copy number quantification is strongly affected by DNA quality and determined a precise correlation betweeMicrobial genomicshttps://www.ebi.ac.uk/biostudies/studies/S-EPMC7660251biostudies-literatureAccurate quantification of bacterial abundance in metagenomic DNAs accounting for variable DNA integrity levels.PMC7660251PRIN 2017Fosso BD'Erchia AMOranger AManzari CPiancone EPesole GfalseAccurate quantification of bacterial abundance in metagenomic DNAs accounting for variable DNA integrity levels.The quantification of the total microbial content in metagenomic samples is critical for investigating the interplay between the microbiome and its host, as well as for assessing the accuracy and precision of the relative microbial composition which can be strongly biased in low microbial biomass samples. In the present study, we demonstrate that digital droplet PCR (ddPCR) can provide accurate quantification of the total copy number of the 16S rRNA gene, the gene usually exploited for assessing total bacterial abundance in metagenomic DNA samples. Notably, using DNA templates with different integrity levels, as measured by the DNA integrity number (DIN), we demonstrated that 16S rRNA copy number quantification is strongly affected by DNA quality and determined a precise correlation betwee2020-01-01T00:00:00Z2020 Oct2026-05-04T19:43:22.329Z2020-11-19T16:59:09ZS-EPMC76602513274995110.1099/mgen.0.000417