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Sureselect human exome library preparation v5 kit

Manufactured by Agilent Technologies
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The SureSelect Human Exome Library Preparation V5 kit is a tool designed for the capture and enrichment of human exonic regions. It provides a comprehensive solution for preparing sequencing libraries from genomic DNA samples.

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Lab products found in correlation

Hiseq x platform, by Illumina (1 mentions) Hiseq 2500 platform, by Agilent Technologies (1 mentions) Truseq dna pcr free library prep kit, by Illumina (1 mentions) Sureselect clinical research exome v1, by Agilent Technologies (1 mentions) Nextseq 500, by Agilent Technologies (1 mentions) Hiseq 2000, by Agilent Technologies (1 mentions) Hiseq x ten, by Illumina (1 mentions) Hiseq 2500, by Illumina (1 mentions) 2100 bioanalyzer high sensitivity dna chip, by Agilent Technologies (1 mentions) Kapa library quantification abi prism kit protocol, by Illumina (1 mentions) Le220 instrument, by Covaris (1 mentions)

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SureSelect kits (5 citations) V5 Sureselect kit (4 citations)

4 protocols using sureselect human exome library preparation v5 kit

1

Whole Genome and Exome Sequencing of Pediatric Patients

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For the genomes of patient (P) P1 and P5, about 1 μg of genomic DNA was submitted to The Centre for Applied Genomics (TCAG, The Hospital for Sick Children); 700 ng DNA was used as input material for library preparation using the Illumina TruSeq PCR-free DNA Library Prep Kit following the manufacturer’s recommended protocol. Validated libraries were then pooled in equimolar quantities and paired-end sequenced on an Illumina HiSeq X platform following Illumina’s recommended protocol to generate paired-end reads of 150 bases in length and about 100 gigabases of raw data per library. For exomes of P1, P2, and P3, good-quality genomic DNA samples were sent to TCAG for exome library preparation using Agilent SureSelect Human Exome Library Preparation V5 kit using the Agilent Bravo Automation System and paired-end sequencing on a HiSeq 2500 platform. For the exomes of P4 and P5, sequencing was performed at The Hospital for Sick Children’s Department of Paediatric Laboratory Medicine using an Illumina NextSeq 500 instrument after enrichment with Agilent SureSelect Clinical Research Exome v1, with 2 × 150 bp reads.
Pasternak Y., Vong L., Merico D., Abrego Fuentes L., Scott O., Sham M., Fraser M., Watts-Dickens A., Willett Pachul J., Kim V.H., Marshall C.R., Scherer S, & Roifman C.M. (2024). Utilization of next-generation sequencing to define the role of heterozygous FOXN1 variants in immunodeficiency. The Journal of Allergy and Clinical Immunology: Global, 3(3), 100267.
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2

Whole Exome Sequencing for Variant Identification

Cited 2 times
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Genomic DNA from the specified PDLs (above) were sent to Genewiz for whole exome sequencing (WES) performed on Illumina HiSeq 2000 (2x150 bp) with libraries prepared using Agilent SureSelect Human Exome Library Preparation V5 kit and sequenced to an average depth of 112.7 (104.9x-119.7x). Raw reads were aligned to the human genome (hg38) using the Burrows Wheeler Aligner (Li and Durbin, 2009 (link)). Variant calling was performed using Mutect 2 ((Cibulskis et al., 2013 (link))) with the following additional filtration steps to retain only high confident mutations: VAF < 0.05, SNV per gene ≥ 3, dbSNPs were all removed.
Hodel K.P., Sun M.J., Ungerleider N., Park V.S., Williams L.G., Bauer D.L., Immethun V.E., Wang J., Suo Z., Lu H., McLachlan J.B, & Pursell Z.F. (2020). POLE mutation spectra are shaped by the mutant allele identity, its abundance and mismatch repair status. Molecular cell, 78(6), 1166-1177.e6.
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3

Whole Genome & Exome Sequencing Protocol

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Next generation sequencing was performed as per the published protocols. Whole genome sequencing (WGS) was performed on an Illumina HiSeq Xten instrument with libraries prepared using the manufacturer’s TruSeq Nano DNA Library Prep kit and sequenced to a depth of 36.1x. For exome sequencing, DNA was enriched using Agilent SureSelect Human Exome Library Preparation V5 kit, then sequenced to a depth of 101.38x (96.61x-108.19x).
Hodel K.P., de Borja R., Henninger E.E., Campbell B.B., Ungerleider N., Light N., Wu T., LeCompte K.G., Goksenin A.Y., Bunnell B.A., Tabori U., Shlien A, & Pursell Z.F. (2018). Explosive mutation accumulation triggered by heterozygous human Pol ε proofreading-deficiency is driven by suppression of mismatch repair. eLife, 7, e32692.
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4

Agilent Exome Sequencing Protocol

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Exome libraries were prepared using Agilent SureSelect Human Exome Library Preparation V5 kit and the Agilent Bravo Automation System followed by paired-end sequencing on an Illumina HiSeq 2500 platform. Genomic DNA (750 ng) was fragmented to 200-bp on average using a Covaris LE220 instrument. Sheared DNA was endrepaired and the 3 0 ends adenylated prior to ligation of adapters with overhang-T. Genomic library was amplified by PCR using 10 cycles and hybridized with biotinylated probes that target exonic regions; the enriched exome libraries were amplified by an additional 8 cycles of PCR. Exomic libraries were validated on a Bioanalyzer 2100 DNA High Sensitivity chip (Agilent Technologies) for size and by qPCR using the Kapa Library Quantification Illumina/ABI Prism Kit protocol (KAPA Biosystems) for quantities. Exome libraries were pooled and sequenced with the TruSeq SBS sequencing chemistry using a V4 high throughput flowcell on a HiSeq 2500 platform following Illumina's recommended protocol. Approximately 6 to 8 gigabases of raw paired end data of 126-bases were generated per exome library.
Stosic A., Fuligni F., Anderson N.D., Davidson S., de Borja R., Acker M., Forte V., Campisi P., Propst E.J., Wolter N.E., Chami R., Mete O., Malkin D., Shlien A, & Wasserman J.D. (2021). Diverse Oncogenic Fusions and Distinct Gene Expression Patterns Define the Genomic Landscape of Pediatric Papillary Thyroid Carcinoma. Cancer research, 81(22).
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