Sequencing of Chicago long-range linkage libraries for chromosome flow sorted chromosome 1H from barley accession Baronesse
Chromosome flow sorting of Baronesse chromosome 1H was performed using the methods described by Dolezel et al. (2012) Functional and Integrative Genomics 12, 397-416 and chromosomal high molecular weight (HMW) DNA was prepared as described in Thind et al. (2017) Nature Biotechnology 35, 793-796. Chicago libraries were generated by in vitro chromatin reconstitution using 250 ng HMW DNA. After fixation with formaldehyde, chromatin was digested with MboI, biotinylated nucleotides were added to 5 prime overhangs, and proximity ligation was performed. After reversal of crosslinking the DNA was treated to remove biotin and sheared to approximately 400 bp. A sequencing library was generated using NEBNext Ultra enzymes (New England Biolabs) and Illumina-compatible adapters.