title: DNA-Seq of Homo sapiens: male and female
design: a total of eight whole blood samples were used for genomic DNA isolation using DNeasy Blood & Tissue kit (Qiagen, Hilden, Germany) following manufacturer's instructions. The DNA quality and quantity were determined using a Nanodrop Lite spectrophotometer (Thermo Scientific, USA) and Qubit Fluorometer 4.0 (ThermoFisher, USA) using the dsDNA high sensitivity method. Whole genome sequencing (WGS) was carried out using DNA nanoball-based technology on the DNBSEQ-G400 (MGI, China) with the appropriate reaction conditions and reagents (MGIEasy Universal DNA Library Prep Set). In brief, the gDNA samples (250 ng) were sheared by enzymatic fragmentation to DNA fragments ranged from 350 to 500 bp, then DNA fragments were selected and purified with MGIEasy DNA Clean Beads (MGI, China). Based on manufacturer's instructions, DNA fragments were end-repaired and dATP-tailed (ERAT) using the appropriate volumes of ERAT buffer and enzyme mix, facilitating the ligation of MGIEasy DNA Adapters. After cleanup adapter-ligated DNA products, a total of 7 cycles of PCR amplification was performed on each sample using using the SimpliAmp Thermal Cycler (Applied Biosystems). Following magnetic bead purification and assessing fragment size distribution using QIAxcel system (QIAxcel DNA high resolution kit), the amplicons were subjected to single-strand circularization processes. After circularization, non-circularized or linear DNA strand was digested with exonuclease and finally obtaining single-strand circular DNA library. The DNA Nanoballs (DNBs) were generated by rolling circle amplification (RCA) technology. The MGIEasy circularization (MGI, China) and  DNBSEQ-G400RS high-throughput sequencing (MGI, China) kits were used for circularized DNA and DNBs, respectively. The DNA nanoballs were loaded onto a sequencing chip and then performed PE100 sequencing on the DNBSEQ-G400 (MGI, China).