Supplementary Data accompanying: Tracking different states of spiked environmental DNA using multiplex digital PCR assays

The Supplementary Data related to the paper Tracking different states of spiked environmental DNA using multiplex digital PCR assays, Environmental Microbiology 2025 contain the following:Sheet 1: Number of target copies measured by dPCR using singleplex and the concomitant multiplex assays. The same dilution series of each target gene was used as DNA template for both singleplex and multiplex assays. Esco_iDNA and Basu_iDNA refer to the cellular spike-ins originating from E. coli and B. subtilis, respectively. Similarly, Esco_exDNA and Basu_exDNA represent the spiked exDNA deriving from each of the model organisms.Sheet 2: Quality control of dPCR assays targeting iDNA spike-ins originating from B. subtilis (∆bglA::kan). Analyses regarding LoD95%, LoQ and linear regression curves are based on these data. Four independent dilution series were created to account for inter-specific assay variation.Sheet 3: Quality control of dPCR assays targeting spiked exDNA originating from B. subtilis (∆licH::kan). Analyses regarding LoD95%, LoQ and linear regression curves are based on these data. Four independent dilution series were created to account for inter-specific assay variation.Sheet 4: Quality control of dPCR assays targeting iDNA spike-ins originating from E. coli (∆chbF::kan). Analyses regarding LoD95%, LoQ and linear regression curves are based on these data. Four independent dilution series were created to account for inter-specific assay variation.Sheet 5: Quality control of dPCR assays targeting spiked exDNA originating from E. coli (∆bglA::kan). Analyses regarding LoD95%, LoQ and linear regression curves are based on these data. Four independent dilution series were created to account for inter-specific assay variation.Sheet 6: Percent recovery of spike-in controls depending on the spiked eDNA states, their bacterial origin and spike-in concentration as well as the environmental samples tested. Esco_iDNA and Basu_iDNA refer to the iDNA spike-ins originating from E. coli and B. subtilis, respectively. Similarly, Esco_exDNA and Basu_exDNA represent the spiked exDNA deriving from each of the model organisms.Sheet 7: Percent recovery of differentially sized DNA fragments used as spike-in controls within selected soil and sludge samples.