GSM8266856: 10_430223-T WES; Homo sapiens; OTHER
Genomic DNA was prepared fosr exome sequencing using the Agilent SureSelect Human All Exon V6 kit with the following steps: Initially, 0.4 μg of high-quality genomic DNA was randomly sheared to a mean size of 180-280 base pairs using a Covaris S220 sonicator. Any remaining DNA overhangs were made blunt by exonuclease and polymerase treatment. Subsequently, the DNA fragments underwent end repair and phosphorylation, followed by the addition of a single 'A' base (A-tailing) and ligation with paired-end adaptors at the 3' ends. Fragments with adaptors on both ends were amplified via PCR. Post-PCR, the library was hybridized with biotinylated probes in solution and then captured using streptavidin-coated magnetic beads to selectively enrich for gene exons. The captured libraries were subjected to an additional PCR to incorporate index tags, preparing them for sequencing. The PCR products were purified with the AMPure XP system, and DNA concentration was determined using a Qubit®3.0 Flurometer. The library size distribution was assessed with NGS3K/Caliper, and the library was quantified by real-time PCR to a final concentration of 3 nM. Ultimately, the DNA library was sequenced on an Illumina platform to achieve paired-end 150 base pair reads.