Sequencing of purified chromosome 3A of T. turgidum cv. Kronos, TILLING line K2282: M5 K2282 mutant line, with yellow flag leaf phenotype
Three batches of 30,000 copies of chromosome 3A derived from M5 K2282 TILLING lines (T. turgidum cv. Kronos) corresponding to ~50 ng of DNA each were sorted into PCR tubes containing 40 l sterile deionized water. Chromosomal DNA was purified and amplified by Illustra GenomiPhi V2 DNA amplification Kit (GE Healthcare, Piscataway, USA) according to (imkov et al., 2008). Library preparation and sequencing were carried out at Novogene. DNA integrity was confirmed on 1% agarose gels. A PCR-free library preparation was carried out, using the NEBNext Ultra II DNA Library Prep Kit for Illumina, following manufacturers instructions. Libraries were sequenced using a HiseqX platform, generating 150 bp paired end reads.