RAD2seq: an efficient protocol for plant genotyping by sequencing

Advances in next-generation sequencing (NGS), through multiplexed sequencing of barcoded samples in a single run, have driven the costs down to the point that is now feasible the re-sequencing of an entire mapping population, also in medium-sized genome species. RAD-seq approach is becoming more popular since implementations of the original protocol have been proposed. A RAD-seq protocol, based on a double-digestion, was recently developed, which reduces sample preparation costs by eliminating random shearing, while increasing the multiplexing power. Here we present a modified two-enzymes RAD-seq (namely RAD2seq) protocol, that includes a biotin/streptavidin guided capturing step which increases the recovering rate of selected fragments, while reducing the cost and time of library preparation.