Genotyping with nextRAD markers allows next-generation sequencing reads to pile up at loci scattered across the genome, giving high-quality SNP calls. It differs from other genotyping-by-sequencing methods by not using restriction enzymes to sample only a portion of the genome. Instead, it uses selective primers to amplify only the loci in the genome with the selective sequence. The small number of steps in the protocol reduces loss during library creation, allowing much lower input compared to other methods.

nextRAD method


Selective primers productively amplify only fragments with the selective sequence at the end. Subsequent sequencing of the library allows determination of genetic variation at these loci.