Human Whole Exome Sequencing
Service Features
● Two exome panels available based on target enrichment with probes: SureSelect Human All Exon v6 (Agilent) and xGen Exome Hybridization Panel v2 (IDT).
- Based on the SureSelect Human All Exon V6 Panel, UTR regions and full-length mtDNA are supplemented to the target capture range.
- UMI-enabled library construction is available with the SureSelect Human All Exon V6 Panel.
● Sequencing on Illumina NovaSeq.
● Bioinformatic pipeline directed towards disease analysis or tumor analysis.
Service Advantages
● Targets Protein Coding Region: By capturing and sequencing protein coding regions, hWES is utilized to reveal variants related to protein structure.
● Cost Effective: hWES yields approximately 85% of the human disease-associated mutations from 1% of the human genome.
● High Accuracy: With high sequencing depth, hWES facilitates the detection of both common variants and rare variants with frequencies lower than 1%.
● Rigorous Quality Control: We implement five core control points across all stages, from sample and library preparation to sequencing and bioinformatics. This meticulous monitoring ensures the delivery of consistently high-quality results.
● Comprehensive Bioinformatics Analysis: our pipeline goes beyond identifying variations to the reference genome, as it incorporates advanced analysis designed to specifically address research questions related to genetic aspects of diseases or tumor analysis.
● Post-Sales Support: Our commitment extends beyond project completion with a 3-month after-sale service period. During this time, we offer project follow-up, troubleshooting assistance, and Q&A sessions to address any queries related to the results.
Sample Specifications
|
Exon Capture Strategy |
Sequencing Strategy |
Recommended Data Output |
|
Agilent SureSelectXT Human All Exon V6 + UTR + mtDNA Agilent SureSelectXT Human All Exon V6 + UTR + mtDNA (with UMI) IDT xGen Exome Hyb Panel V2 |
Illumina NovaSeq PE150 |
5 -10 Gb For mendelian disorders/rare diseases: > 50x For tumor samples: ≥ 100x |
Sample Requirements
|
Sample Type |
Amount (Qubit® ) |
Concentration
|
Volume |
Agarose Gel |
|
WES-FF |
≥ 150 ng | ≥ 15 ng/µL |
≥ 15 µL |
No or limited contamination and degradation (The main band is clear). FFPE DNA: If the main band is unclear, a distinct smear above 2 kb is required. |
|
WES-FFPE |
≥ 200 ng | ≥ 15 ng/µL |
Bioinformatics
Bioinformatic analysis of hWES-disease samples includes:
● Sequencing data QC
● Reference Genome Alignment
● Identification of SNPs and InDels
● Functional Annotation of SNPs and InDels
Bioinformatic analysis of tumor samples includes:
● Sequencing data QC
● Reference Genome Alignment
● Identification of SNPs, InDels and somatic variations
● Identification of germline variants
● Mutation signatures analysis
● Identification of drive genes based on gain-of-function mutations
● Mutation annotation at the level of drug susceptibility
● Heterogeneity analysis – calculation of purity and ploidy
Service Work Flow
Sample delivery
DNA extraction
Library construction
Sequencing
Data analysis
Data delivery
After-sale services
Data QC – Statistics of Exome capture
Variant identification – InDels
Advanced analysis: identification and distribution of deleterious SNPs/InDels – Circos plot
Tumor analysis: identification and distribution of somatic mutations – Circos plot
Tumor analysis: clonal lineages
