Bar32
Acid-delinted seeds of B713 and Bar32-30 (G. hirsutum) were chipped and rolled in moist paper towels. Rolled towels were placed in an incubator set at 30 C for 4 days to germinate. DNA was extracted using the Qiagen Genomic Tip kit (Qiagen Inc.). The sequencing libraries were constructed at the BYU DNA Sequencing Center (DNASC). DNA shearing of both libraries was done on a Megaruptor2 (~20 kb). HMW DNA was partitioned into 13 bin using the Sage Elf and the top 5 bins were run on a Fragment Analyzer (Agilent Inc.) to select the appropriate bin size range (15-18 kb). Libraries were made using the SMRTbell Express Template Prep kit as recommended by PacificBiosciences. Five PacBio cells were sequencing from each library on the Pacific Biosciences Sequel 2 system. The PacBio reads were assembled using hifiasm (Cheng et al. 2021) using default parameters. Both assembled genomes were aligned to previously assembled genomes of G. hirsutum using minimap2 (Li 2018) and visualized by dotPlotly (https://github.com/tpoorten/dotPlotly). Manual scaffolding was used to create pseudomolecules of both genomes.