WES library construction and the sequencing were performed by Biomed lab company (Shanghai, China). In brief, WES libraries were generated by TruSeq DNA Exome Kit according to the manufacturer’s protocol, and the Exome was captured by SureSelect Human All Exon V5 or V6 kit (Agilent Technologies, Inc., United States). Illumina Hiseq (2) and (500) platform was deployed to sequence the shotgun libraries, and paired-end (PE) reads were generated (150 nt for AVSD, 125 nt for control).
Sureselect human all exon v5 or v6 kit
Manufactured by Agilent Technologies
Sourced in United States
The SureSelect Human All Exon V5 or V6 kit is a targeted enrichment solution for whole exome sequencing. It is designed to capture the protein-coding regions of the human genome, known as the exome. The kit includes a set of RNA baits that hybridize to the exonic regions, allowing for the selective isolation and sequencing of these regions.
Automatically generated - may contain errors
3 protocols using sureselect human all exon v5 or v6 kit
1
Whole Exome Sequencing Library Prep and Sequencing
2
Whole Exome Sequencing from Diverse Biosamples
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WES was performed in 55 baseline biological samples available at VHIO’s sample biobank (19 germline DNA, 22 tumor DNA, 5 patient-derived xenograft DNA and 9 plasma cfDNA) from 28 patients from the discovery cohort. Genomic libraries were prepared using 10–15 ng of cfDNA and the ThruPLEX DNA or Plasma-seq Kit (Rubicon Genomics; now commercialized as the SMARTer ThruPLEX plasma-seq kit by Takara Bio)37 (link),38 (link). Quality control of libraries was carried out in TapeStation (Agilent), and amplified profiles of ~300 base pairs were considered for downstream analysis. Capture of the genomic coding regions for WES was performed using the SureSelect Human All Exon V5 or V6 kit (Agilent), with a target sequencing output of 12 Gb (100×) for genomic DNA and 36 Gb (300×) for tumor and cfDNA. Sequencing was carried out on a HiSeq or NovaSeq Illumina sequencing platform.
3
Targeted Exome Sequencing of Blood and Muscle
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We performed ES and Sanger sequencing as described in our previous publication. 18 DNA was extracted from patients' blood leukocytes with the QuickGene-610L system (Tokyo, Japan, Fujifilm) following the manufacturer's instructions, or from frozen skeletal muscle using a conventional phenol chloroform extraction method. DNA was captured with the SureSelect Human All Exon V5 or V6 Kit (Agilent).
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