Method Category: Genome > Rearrangements and Markers
Description: Double digest restriction-site associated DNA (ddRADseq), also called ddRAD, is a variation on the RAD sequencing protocol, which is used for SNP discovery and genotyping. , In this variation, the fragment shearing is replaced with a second restriction digestion to improve the tunability and accuracy of the size-selection step. The protocol also includes a second index to allow combinatorial indexing. Several RAD variations 2b-RAD, SLAF-seq, and hyRAD have been developed to address specific applications and there are multiple software packages available to analyze RAD data.
In this method, genomic DNA is first digested with a restriction enzyme and a barcoded P1 adaptor ligated to the fragments. The adaptor-ligated fragments from different samples are combined, if samples are multiplexed, and the DNA digested by a second restriction enzyme. The fragments are size-selected and purified. The P2 primers are then ligated and the fragments amplified.
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