GENOTYPING BY NEXT-GENERATION SEQUENCING

摘要

Provided herein is technology relating to genotyping and particularly, but not exclusively, to methods for genotyping one or more organisms by genome sequencing. In some embodiments of the technology, two restriction enzymes are used to generate two sites with different overhangs at each end of the digested fragments. One site is ligated with the barcode oligonucleotide to permit multiplexing of samples during analysis, e.g., sequencing. The other site is ligated with an oligonucleotide without the barcode. The number of sites targeted for analysis is further reduced by the design and selection of amplification primers complementary to the non-barcode site.