State-Specific Extraction of Environmental DNA: Spike-and-Recovery Controls to Validate and Optimise Extraction Protocols.

ABSTRACT: Getting insights into the quantitative and qualitative contribution of different DNA states, i.e., extracellular (exDNA) and intracellular DNA (iDNA), to the total environmental DNA (eDNA) pool requires reliable methods for their separation. Even though a multitude of respective extraction protocols has been published, their validation is often missing. Here, we selected four protocols for the state-specific extraction of eDNA and traced the separation of exDNA and iDNA within natural environments using previously designed spike-and-recovery controls. Besides accounting for the different eDNA states, the spike-ins also distinguished different bacterial origins (gram-positive, gram-negative). Following their quantification by digital PCR, the recovery of exDNA and iDNA spike-ins in both the

SUBMITTER: Zohrer J 

PROVIDER: S-EPMC12672054 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature