title: WES of homo sapiens: adult male blood
design: Total DNA was extracted from the patients peripheral blood samples. DNA was broken into 200300 bp lengths after passing DNA quality control, and specific linker sequences with tag sequences (Index) were added to both ends of the fragmented DNA. A DNA library was obtained following polymerase chain reaction (PCR) amplification. DNA library with double-stranded DNA was denatured to single-stranded DNA (ssDNA) using high temperature, DNA ligase, and circularization primers to link the two ends of the ssDNA to form a loop. The non-circularized ssDNA in the reaction system was digested and degraded to obtain circular ssDNA that can be used for preparing DNA nanoballs. The prepared DNA Nano Ball was then loaded onto a Patterned Array and sequenced using Combinatorial Probe-Anchor Synthesis. After the polymerization reaction of the sequencing primer-anchored molecules and fluorescent probes on the DNA Nano Ball, the optical signal was captured, read, and identified using a high-resolution imaging system to obtain single base sequence information. The next cycle was performed to obtain the next base sequence information. Sequencing results were bioinformatically analyzed, and variant loci were annotated with databases related to variant frequency, protein hazard, and disease.