CGCG9
Initial DNA sample quality assessment, DNA library preparations and sequencing were conducted at GENEWIZ (Leipzig, Germany) as follows: The genomic DNA was quantified using the Qubit 4.0 Fluorometer and qualified using the Agilent 5300 Fragment Analyzer. Enrichment probes were designed against the region of interest and synthesized through Agilent Technologies (Santa Clara, CA, USA). Library preparation was performed according to the manufacturers guidelines. Briefly, the genomic DNA was fragmented by acoustic shearing with a Covaris E220 instrument. Fragmented DNA was end repaired and adenylated at the 3ends. Adapters were ligated to the DNA fragments, and adapter-ligated DNA fragments were enriched with limited cycle PCR. Adapter-ligated DNA fragments were validated using Agilent Fragment Analyzer (Agilent Technologies, Santa Clara, CA, USA), and quantified using Qubit 2.0 Fluorometer. Adapterligated DNA fragments were hybridized with biotinylated baits. The hybrid DNA was captured by streptavidin-coated binding beads. After extensive wash, the captured DNA was amplified and indexed with Illumina indexing primers. Post-captured DNA libraries were validated using 5300 Fragment Analyzer (Agilent, Santa Clara, CA, USA) and quantified using Qubit 2.0 Fluorometer. Illumina reagents and kits for DNA library sequencing cluster generation and sequencing were used for enrichment DNA sequencing. Post-captured libraries have been multiplexed on a flowcell and loaded on the Illumina NovaSeq 6000 instrument according to the manufacturer's instructions. The samples were sequenced using a 2x150 paired-end (PE) configuration. Image analysis and base calling was conducted by the NovaSeq Control Software on the NovaSeq instrument. Raw sequencing data (.bcl files) generated from Illumina NovaSeq were converted into fastq files and de-multiplexed using Illumina's bcl2fastq software.