Genomic DNA Extraction and GBS Library Preparation

Total genomic DNA was extracted from muscular tissues for each sample using the Qiagen^® DNeasy Blood and Tissue extraction kit (QIAGEN, Valencia, CA, USA) according to the manufacturer’s protocol. Following the protocol of Elshire et al. [35 (link)], we generated the GBS libraries. First, genomic DNA was digested using MseI restriction enzymes (New England Biolabs, NEB, Ipswich, MA, USA) at 37 °C. Next, after adding a sequencing junction at the end of the enzyme digestion, the purified samples were subjected to PCR reactions using a Gel Extraction Kit (QIAGEN, Valencia, CA, USA), and fragments retaining approximately 300–350 bp were isolated from agarose gels. Moreover, using a 150 bp paired-end protocol, the obtained library was sequenced on the Illumina NovaSeq6000 platform at Novogene Bioinformatics Technology Co., Ltd., Beijing, China (www.novogene.cn; accessed on 15 June 2022). Finally, the original sequences (sequencing reads) are obtained, which we call raw data.