show Abstracthide AbstractPCR-free libraries, constructed from genomic DNA extracted from young leaf tissue, were sequenced using Illumina NovaSeq 6000 using paired-end 150. The collection contains 328 genotypes belonging mostly to the Aegilops and Triticum genera. All 11 diploid, 10 tetraploid and 4 hexaploid Aegilops species are represented by 2 to 12 accessions each. Two diploid, two tetraploid and two hexaploid Triticum species are represented. In addition, one accession each of Haynaldia villosa, Thinopyrum bessarabicum and Th. intermedium are also represented. The 92 diploid accessions were sequenced using each ~1%, the 140 tetraploid ones using ~2% and the 96 hexaploid ones using ~3% of a NovaSeq lane to achieve ~1-2X coverage across the 328 accessions onto a total of 7 Novaseq 6000 lanes.