A Robust, Simple Genotyping-by-Sequencing (GBS) Approach for High Diversity Species (original) (raw)
Figure 2
Steps in GBS library construction.
Note: Up to 96 DNA samples can be processed simultaneously. (1) DNA samples, barcode, and common adapter pairs are plated and dried; (2–3) samples are then digested with _Ape_KI and adapters are ligated to the ends of genomic DNA fragments; (4) T4 ligase is inactivated by heating and an aliquot of each sample is pooled and applied to a size exclusion column to remove unreacted adapters; (5) appropriate primers with binding sites on the ligated adapters are added and PCR is performed to increase the fragment pool; (6–7) PCR products are cleaned up and fragment sizes of the resulting library are checked on a DNA analyzer(BioRad Experion® or similar instrument). Libraries without adapter dimers are retained for DNA sequencing.