Tracking Different States of Spiked Environmental DNA Using Multiplex Digital PCR Assays.

ABSTRACT: The study of microbial communities based on the total environmental DNA (eDNA) is influenced by the presence of different eDNA states, i.e., intracellular (iDNA) and extracellular DNA (exDNA), and the choice of the DNA extraction method. Although the use of spike-and-recovery controls facilitates the diagnosis of such issues, appropriate experimental setups simultaneously accounting for the different eDNA states and their bacterial origins are missing. Here, we used two single-gene deletion mutants of both Escherichia coli and Bacillus subtilis to trace exDNA and iDNA spike-ins of each selected model organism within environmental samples. Unique primer/probe sets were developed for each strain, allowing their absolute quantification using multiplex digital PCR assays. The proposed spike-an

SUBMITTER: Zohrer J 

PROVIDER: S-EPMC11950903 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature