Abstract

Quantification of Bacterial DNA for Metagenomic Control Materials

Denise O'Sullivan, Researcher, LGC Genomics

Advances in DNA sequencing technology enable the analysis of the genetic content of microbial populations (the metagenome) as a whole. Associated studies commonly measure the different quantities of the component organisms. Early evidence suggests however that sample processing and preparation methods can have a strong influence on the measured abundance of the respective microbes. In this study we use digital PCR to quantify the component organisms of a metagenomic control material comprising 10 common human pathogens mixed together in different proportions. We investigate how targeting different bacterial chromosomal positions effect the measurement, compare dPCR with fluorescence and absorbance based methods as well as next generation sequencing approaches. We demonstrate how dPCR may offer a simple method for providing confidence when measuring complex metagenomes.