GCN sample analysis via single species assay and metabarcoding (NECR534)

The aim of this project was to compare the results of great crested newt (GCN) environmental DNA (eDNA) analysis from samples collected using either ethanol precipitation or filtration methods and to investigate whether GCN could be reliably detected from a sub-set of the samples collected via filtration methods using DNA metabarcoding. 100 samples were collected using the standard ethanol precipitation-based sampling kits and during the same sampling visit a sample was also taken using a Sterivex-HV Pressure Filter Unit with a 0.45µm pore size. GCN analysis was performed using the methods in WC1067 and via metabarcoding of a sub-set of 20 of the samples all of which were positive for GCN with both sampling methods. GCN species-specific results comparison of the two sample collection methods were largely in agreement with 27 of the samples being positive for GCN eDNA by both methods with 5 additional positive samples for both the ethanol precipitation and filtration collected samples respectively. Metabarcoding of 12S DNA for vertebrate species resulted in 29 species being detected across the 20 samples with 11 of the 20 samples detecting great crested newt DNA.