RS12_bleached
For each sample, PCRs were performed in triplicate using the Qiagen Multiplex PCR kit, between 1 and 35 ng template DNA, a primer concentration of 0.2 ?, and a final reaction volume of 25 ?. PCRs were performed as follows: One cycle at 95 ? for 15 min, 30 cycles each at 95 ? for 30 s, 55 ? for 90 s, and 72 ? for 30 s, followed by a final extension step at 72 ? for 10 min. Triplicate PCRs for each sample were pooled and cleaned with the Agencourt AMPure XP magnetic bead system (Beckman Coulter, Brea, CA, USA), and subsequently underwent an indexing PCR to add Nextera XT barcoded sequencing adapters (Illumina) according to the manufacturer? instructions.?he library was sequenced on the Illumina MiSeq, 2*300 bp paired-end version 3 chemistry according to the manufacturer? specifications.
SUB2118529
RS12_bleached