HiPlex amplicon sequencing of Triticum aestivum, Secale cereale, and x Triticosecale
Total genomic DNA was extracted from fresh leaf material using the E.Z.N.A. Plant DNA DS Mini Kit (Omega Bio-tek; Norcross, USA). DNA samples were quantified using Quantus and concentrations were normalized (range 10-20 ng/l). Amplicons for 451 loci were amplified in a multiplex PCR per sample, while adding sample-specific indices (Floodlight Genomics, USA). Illumina sequencing adapters were ligated with the Kapa HiFi PCR-free ligation kit, and libraries were paired-end sequenced on an Illumina NovaSeqX instrument (Admera Health, USA). All read data were demultiplexed, barcodes and adapter sequences were removed, and forward and reverse reads were merged to a single read.
SUB16031324
K0020_HiPlex_rep1