Reduced representation sequencing of pooled DNA from 88 Secale cereale accessions
10l of the normalized DNA from each accession was double-digested with PstI and MspI restriction enzymes then ligated to two adapters with barcode. After the adapters were ligated, the samples were pooled together for PCR amplification and subsequently size-selected for 250-300bp amplicons using the E-gel system . Libraries (80 pM) were prepared and sequenced in an Ion Proton System using Ion PI Hi-Q Chef and Sequencing 200 kits with two Ion P1v3 chips .