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- Title
Use of RNase H‐dependent PCR for discrimination and detection of closely related species from environmental DNA.
- Authors
Rodgers, Torrey W.; Olson, John R.; Mock, Karen E.; Jarman, Simon
- Abstract
Species‐specific, probe‐based quantitative PCR (qPCR) assays are now commonly used to detect aquatic species from environmental DNA. However, probe‐based qPCR alone does not always provide the specificity needed to distinguish closely related, congeneric species, which may result in amplification of non‐target DNA, causing false positives from eDNA samples.Here, we developed species‐specific qPCR assays using RNase H‐dependent PCR (rhPCR) for detecting closely related fish species from environmental DNA.We found that rhPCR allowed us to achieve specificity that was not possible with TaqMan®qPCR alone, and we used these assays for species detection from eDNA samples.Use of rhPCR will allow species‐specific detection from environmental DNA for a broad range of species including those that occur in sympatry with other closely related, congeneric species, which has not always been possible with probe‐based qPCR alone.
- Subjects
DNA; GENE amplification; SPECIES; DISCRIMINATION (Sociology)
- Publication
Methods in Ecology & Evolution, 2019, Vol 10, Issue 7, p1091
- ISSN
2041-210X
- Publication type
Article
- DOI
10.1111/2041-210X.13187