Trusight oncology 500 library preparation kit

Manufactured by Illumina

Sourced in United States

The TruSight Oncology 500 Library Preparation Kit is a laboratory equipment product that enables the preparation of nucleic acid libraries for targeted DNA sequencing. The kit provides reagents and protocols to generate libraries from genomic DNA samples for analysis on Illumina sequencing platforms.

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Lab products found in correlation

Qubit high sensitivity kit, by Thermo Fisher Scientific (1 mentions) Nextseq 2000 system, by Illumina (1 mentions) Trusight oncology 500 kit, by Illumina (1 mentions) Nexseq550, by Illumina (1 mentions) Generead dna ffpe kit, by Qiagen (1 mentions) Microtube 50 afa fiber screw cap, by Covaris (1 mentions) M220 focused ultrasonicator, by Covaris (1 mentions) Qci interpret, by Qiagen (1 mentions) Nextseqtm 550dx, by Illumina (1 mentions) Trusight oncology 500, by Illumina (1 mentions) Novaseq 6000, by Illumina (1 mentions)

4 protocols using trusight oncology 500 library preparation kit

Profiling KRAS G12 Variants in cfDNA

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The analyses of KRAS G12 variants in cfDNA were performed via NGS using the NextSeq 2000 System (Illumina, CA, USA). Approximately 40 ng cfDNA per sample was used for library preparation using the Trusight Oncology 500 Kit (Illumina) according to the manufacturer’s instructions. The sequence data were processed and analyzed using the TruSight Oncology 500 Local App version 1.3 (Illumina). The average sequencing depth was approximately 1500×. Variant allele frequencies (VAFs) were calculated as altered variant reads/total reads.
Genomic DNA was extracted from 6 × 10 μm tissue sections using the GeneRead DNA FFPE Kit (Qiagen, Venlo, The Netherlands). DNA concentrations were measured using a Qubit high-sensitivity kit (Thermo Fisher Scientific). Subsequently, 40 ng of DNA was used as input for library preparation. Input DNA was sheared on a Covaris M220 Focused-ultrasonicator (Covaris, MA, USA) using a microTUBE-50 AFA Fiber Screw-Cap (Covaris) following the manufacturer’s instructions. DNA libraries were prepared using the hybrid capture-based TruSight Oncology 500 Library Preparation Kit (Illumina) following Illumina’s assay protocol. Libraries were sequenced using NexSeq550 (Illumina).

Huang C.J., Huang W.Y., Chen C.Y., Chao Y.J., Chiang N.J, & Shan Y.S. (2022). Cancer-cell-derived cell-free DNA can predict distant metastasis earlier in pancreatic cancer: a prospective cohort study. Therapeutic Advances in Medical Oncology, 14, 17588359221106558.

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Hybrid Capture-based Tumor Sequencing

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A selection of tumor samples was subjected to sequencing. Library preparation was performed using the hybrid capture-based TruSight Oncology 500 Library Preparation Kit (TSO500; Illumina) following the manufacturer’s protocol. DNA preparation and sequencing were performed as described before.45 (link) 40–100 ng DNA was used as input for the library preparation, depending on the age and quality of the DNA sample.

Gorris M.A., van der Woude L.L., Kroeze L.I., Bol K., Verrijp K., Amir A.L., Meek J., Textor J., Figdor C.G, & de Vries I.J. (2022). Paired primary and metastatic lesions of patients with ipilimumab-treated melanoma: high variation in lymphocyte infiltration and HLA-ABC expression whereas tumor mutational load is similar and correlates with clinical outcome. Journal for Immunotherapy of Cancer, 10(5), e004329.

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Targeted Sequencing of Oncology Genes

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The hybrid capture-based TruSight Oncology 500 Library Preparation Kit (Illumina, San Diego, CA, USA) was employed for library preparation according to the manufacturer’s protocol. Unique molecular identifier (UMI)-containing adaptors were ligated to the DNA fragments. Next, a pool of oligos specific to the 523 genes included in TSO500 were hybridized to the DNA libraries. Streptavidin magnetic beads captured the probes hybridized to the targeted regions. Primers were used to amplify the enriched libraries before purification with sample purification beads. Afterward, the libraries were pooled, denatured, and diluted. TSO500 libraries were sequenced on a NextSeq^TM 550Dx (Illumina). Variant annotation was performed using Qiagen Clinical Insight (QCI) Interpret (Qiagen N.V., Venlo, The Netherlands). In addition, the software vcf2maf converter (https://github.com/mskcc/vcf2maf; accessed 1 June 2022) was employed to generate maf files, which were uploaded to a locally installed version of cBioportal [32 (link),33 (link)] for analysis of patient and cell line data.

Scheiter A., Hierl F., Winkel I., Keil F., Klier-Richter M., Coulouarn C., Lüke F., Kandulski A., Evert M., Dietmaier W., Calvisi D.F, & Utpatel K. (2022). Wnt/β-Catenin-Pathway Alterations and Homologous Recombination Deficiency in Cholangiocarcinoma Cell Lines and Clinical Samples: Towards Specific Vulnerabilities. Journal of Personalized Medicine, 12(8), 1270.

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Targeted Sequencing of Cancer Genes

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Samples with a sufficient quantity of DNA (minimum input DNA was set to 250 ng) was available in 33 out of the 76 cases. Library preparation was performed using the hybrid-capture TruSight Oncology 500 Library Preparation Kit (Illumina), targeting 523 frequently altered genes in cancer (Supplementary Material). We followed the TruSight Oncology 500 Reference Guide from Illumina (Document no: 1000000067621; https://support.illumina.com/downloads/trusight-oncology-500-reference-guide-1000000067621.html). Libraries were sequenced on a NovaSeq 6000 (Illumina) for 2 × 150-bp paired-end reads according to the manufacturer's protocol.

Ramberg I., Vieira F.G., Toft P.B., von Buchwald C., Funding M., Nielsen F.C, & Heegaard S. (2021). Genomic Alterations in Human Papillomavirus–Positive and –Negative Conjunctival Squamous Cell Carcinomas. Investigative Ophthalmology & Visual Science, 62(14), 11.

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