RAD-seq of Pacific Oyster: Molluscan Broodstock Program, Oregon State University
DNA was extracted from adductor muscle sections of Crassostrea gigas, purified and quantified by fluorometry, then normalized to 10 ng/ul and used as an input to double-digest restriction site associated (RAD)-sequencing library preparation. Genomic DNA was digested using NsiI and MspI cutting enzymes, and size selected. Samples were individually barcoded and multiplexed at 32 individuals per chip. Multiplexed libraries were sequenced on an Ion Torrent Proton. Fastq data were trimmed for adapters and low quality sequences, then demultiplexed and truncated to a consistent size (80 bp), then uploaded to NCBI.