Barkley Canyon sedaDNA V9 metabarcoding
V9 region of 18S rRNA gene amplified with 1389F - 1510R tag-encoded primers (i.e. unique 8-nt sequence added to 5' end of each primer) with 38 cycles, purified with the Qiagen MinElute PCR Purification Kit (Qiagen, Cat.No. 28004), triplicates PCR reactions pooled and again equimolar pooled into two libraries (1A & 1B), and prepared for sequencing following the TruSeq DNA PCR-Free Library Preparation Kit (Illumina, Cat. No. 20015962) prior to sequencing on MiSeq using a V2 Micro 300 cycle kit (2 x 150bp).