sierras-bumble-bee-rad-seq-for-sibship-study
RAD libraries were produced by transferring 10 l of DNA to a new plate and cleaving with PstI (New England Biolabs [NEB]). Unique barcodes were then attached to the cleaved DNAs via a biotinylated adapter. Samples were then pooled for each plate, and the pooled DNA was sheared and purified with magnetic streptavidin beads. SbfI-HF (NEB) was used to remove DNA from the beads. Each plate library received a unique barcode using a NEBNext Ultra DNA Library Prep Kit for Illumina. Lastly, libraries from each plate were pooled in an equimolar ratio and sequenced at the UC Davis Genome Center on an Illumina HiSeq 4000 using paired end 150 bp reads.