Project name: Hordeum vulgare subsp. vulgare cultivar:Baronesse

Description: Our goal is to resolve the Mla3 locus on chromosome 1H from the 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 Dovetail sequencing of the chromosome was performed by Dovetail Genomics (Santa Cruz, CA, USA), with initial assembly in Meraculous and final scaffolding in HiRise. Briefly, 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 and sequenced using Illumina HighSeq X using two insert sizes, 221 bp and 454 bp, with 266.3 and 246.9 million paired end reads, respectively. An initial assembly using Meraculous had length 450.7 Mb on 40,855 scaffolds. The HiRise assembly had 454.5 Mb on 2,009 scaffolds.

Data type: Genome sequencing and assembly

Sample scope: Monoisolate

Relevance: Agricultural

Release date: 2022-11-10

Last updated: 2022-09-12