Reseq of Gossypium hirsutum: mature wild leaf
The dry cotton leaf tissue (about 30 to 50 mg) was disrupted into fine powders with 2.0-mm ZR Bashing Beads (Zymo Research, Irvine, CA, USA) using the Geno/Grinder 2010 system (SPEX SamplePrep, Metuchen, NJ, USA) at 1500 rpm for 30 sec. The genomic DNA was extracted from cotton leaf powder using Qiagen Plant DNeasy Mini kit (Qiagen, Germantown, MD, USA) following the manufacturers instruction. The concentration and purity of extracted genomic DNA were measured by the NanoDrop One spectrophotometer (Thermofisher Scientific, Waltham, MA, USA). The genomic DNA integrity was validated by agarose gel electrophoresis. A total of 300 mg to 500 mg of genomic DNA from each cotton sample was subjected to the Illumina DNA-Seq library preparation using NEBNext Ultra II FS DNA Library Prep Kit and NEBNext Multiplex Dual Index Primers Set (New England Biolabs, Ipswich, MA, USA) based on the manufacturers manuals. Genomic libraries were equimolarly pooled together and sequenced on one lane of NovaSeq S4 system (Illumina, San Diego, CA) with paired-ended 150 bp sequencing length by Novogene Corporation (https://www.novogene.com/us-en/).