Multiplexed Shotgun Genotyping for Rapid and Efficient Genetic Mapping
10 ng of genomic DNA from each individual was digested with the restriction enzyme MseI and 96 custom-designed adapters, each carrying a unique 6 bp barcode, were ligated to the restriction fragments. The ligation reactions were combined, fragments were size-selected, and standard Illumina flow-cell adaptors were added using PCR with custom designed primers. Sequencing this library produced approximately 22 million 101 bp reads in a single lane of an Illumina Genome Analyser.