Our highest-performing human whole-exome sequencing solution, delivering focused and comprehensive exome enrichment.
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Hands-on time
Input quantity
Illumina DNA Prep with Exome 2.5 Enrichment v2 kits offer a fast, high performing, and cost-effective workflow for reliable human whole-exome sequencing (WES).
Easily add targets to your WES experiment, or boost coverage for critical targets, with a supplementary custom enrichment panel. Add your customized Illumina Custom Enrichment Panel v2 as a spike-in to Illumina DNA Prep with Exome 2.5 Enrichment v2 hybridization reactions. Design your customized WES spike-in panel using the Illumina DesignStudio tool, a free sequencing assay design tool, or with help from the Illumina Concierge design team. Additional mitochondrial genome coverage can be incorporated using the Twist Bioscience for Illumina Mitochondrial Panel, either independently or in combination with a custom spike-in panel.
Kit includes library preparation and hybridization reagents, the Twist Bioscience for Illumina Exome 2.5 Panel, cleanup and size-selection beads, and indexing components. Complete your WES workflow by sourcing library prep, sequencing, and DRAGEN analysis directly from Illumina.
| Assay time | 6.5 hr |
|---|---|
| Automation capability | Liquid handling robot(s) |
| Content specifications |
Twist Bioscience for Illumina Exome 2.5 Exome 2.5 Panel 37.5 Mb coding content (≥ 99% of RefSeq, CCDS, ClinVar and ACMG pathogenic/likely pathogenic variants, COSMIC Cancer Gene Census), OMIM Twist Bioscience for Illumina Mitochondrial Panel covers the circular mitochondrial genome ChrM (16,659 bp, 37 genes) |
| Hands-on time | ~2 hr |
| Input quantity | 50–1000 ng DNA |
| Instruments | NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NextSeq 550Dx in Research Mode, NovaSeq X System, NovaSeq 6000Dx in Research Mode, NovaSeq 6000 System, NovaSeq X Plus System |
| Mechanism of action | Bead-linked transposome and hybrid-capture chemistry |
| Method | Targeted DNA sequencing, Exome sequencing, Target enrichment |
| Multiplexing | Pre-enrichment pooling of up to 12-plex is tested. Kit configuration supports multiplexing up to 192 samples, up to 384 samples is possible with added unique dual index sets. |
| Nucleic acid type | DNA |
| On-target reads | >90% based on performance during internal testing |
| Specialized sample types | Blood, Saliva |
| Species category | Human |
| Technology | Sequencing |
| Uniformity | >97% based on average performance from internal testing on control gDNA samples |
| Variant class | Single nucleotide polymorphisms (SNPs), Loss of heterozygosity (LOH), Germline variants, Single nucleotide variants (SNVs), Insertions-deletions (indels), Copy number variants (CNVs) |
Illumina DNA Prep with Exome 2.5 Enrichment v2 kits include library prep, enrichment, enrichment probe panel, purification beads, and index adapter reagents. Choose a different index set if preferred.
The Twist Bioscience for Illumina Mitochondrial Panel is an optional fixed content spike-in panel that allows enrichment and sequencing of mitochondrial DNA variants.
The Illumina Custom Enrichment v2 probe panels are optional customizable enrichment panels which can be added to the exome workflow as a spike-in to extend or boost target coverage.
For direct blood input, the Flex Lysis Reagent Kit is required.
The Infinium MIDI Heatblock Insert fits in the SciGene HybEx Microheating system and is required for the enrichment step.
Illumina DNA Prep with Exome 2.5 Enrichment v2 kits contain the Twist Bioscience for Illumina Exome 2.5 panel. Optional panels may be added to the hybridization reaction to boost or extend coverage.
Twist Bioscience for Illumina Mitochondrial Panel or Illumina Custom Enrichment Panel v2 may be used as spike-in panels. The user guide supports the use of up to two spike-in panels in the exome hybridization reaction.
Custom enrichment panels can be designed through the DesignStudio design tool or Illumina Concierge design team.
Illumina DNA Prep with Exome 2.5 Enrichment v2 is part of a comprehensive workflow that supports whole-exome sequencing from sample preparation through data analysis and interpretation.
Illumina DNA Prep with Exome 2.5 Enrichment v2
Not available in all countries. Illumina Connected Insights supports user-defined tertiary analysis through API calls to third-party knowledge sources.
| Instrument | Recommended number of samples | Read length |
|---|---|---|
| NextSeq 2000 System | Samples per run (by flow cell type): P2: 19, P3: 57 (based on 50× mean target coverage, 42M reads, 4.2 Gb of data) |
2 × 101 bp |
| NovaSeq 6000 System | Samples per run by flow cell type: SP: 34, S1: 69, S2: 176, S4: 428 based on 50× mean target coverage, 42M reads, 4.2 Gb of data. Note: S4: 428 requires NovaSeq Xp workflow or additional indexes. |
2 × 101 bp |
Whole-exome sequencing is used to investigate protein-coding regions of the genome to uncover genetic influences on disease and population health.
Genetic and complex disease research
Comprehensive array and next-generation sequencing solutions to accelerate research of various genetic complex diseases.
Our sequencing and microarray technologies support a broad range of cancer genomics research applications, from DNA to RNA analysis, epigenetics, and more.
Improved performance of Illumina DNA Prep with Exome 2.5 Enrichment v2
Libraries were constructed and enriched using either the Illumina DNA Prep with Exome 2.5 Enrichment or Illumina DNA Prep with Exome 2.5 Enrichment v2. Enriched libraries were sequenced on a NovaSeq 6000 System and analyzed using DRAGEN Enrichment App.
Consistent coverage of disease-associated targets
Illumina DNA Prep with Exome 2.5 Enrichment v2 using a 1.5-hr (orange) or 16-hr (blue) hybridization time shows high coverage at ≥ 20× of targets from public databases.
Performance metrics
Illumina DNA Prep with Exome 2.5 Enrichment v2 shows high enrichment and coverage metrics at both 1.5 hr (orange) and 16 hr (blue) hybridization time.
Increased coverage of select targets with custom spike-in panel
The Illumina DNA Prep with Exome 2.5 Enrichment v2 protocol was performed with and without a custom spike-in panel targeting regions covered by the Twist Bioscience for Illumina Exome 2.5 Panel. Coverage was then assessed across the regions targeted by the spike-in panel.
Illumina DNA Prep with Exome 2.5 Enrichment v2 - A scientist's perspective
Exome uniformity matters more than you think. Massimo Delledonne, Professor of Genetics at University of Verona, Italy, shares hands-on insights from evaluating Illumina DNA Prep with Exome 2.5 Enrichment v2. He talks about why exome uniformity, reproducibility, and workflow simplicity matter for real‑world research.
Illumina DNA Prep with Exome 2.5 Enrichment v2, (S) Tagmentation Set B (96 Samples, 12-plex)
20155541
Includes Illumina DNA Prep with Enrichment library preparation and enrichment reagents, Twist Bioscience for Illumina Exome 2.5 Panel, Illumina Purification Beads, and Illumina DNA/RNA UD Indexes Set B, Tagmentation for whole exome enrichment of 96 libraries (eight, 12-plex enrichment reactions).
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Illumina DNA Prep with Exome 2.5 Enrichment v2, (S) Tagmentation Set D (96 Samples, 12-plex)
20155543
Includes Illumina DNA Prep with Enrichment library preparation and enrichment reagents, Twist Bioscience for Illumina Exome 2.5 Panel, Illumina Purification Beads, and Illumina DNA/RNA UD Indexes Set D, Tagmentation for whole exome enrichment of 96 libraries (eight, 12-plex enrichment reactions).
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Yes, gDNA input of 10–49 ng can be used. However, this protocol does not normalize final pre-enrichment library yields < 50 ng and, therefore, quantification and normalization of libraries before and after enrichment is required.
For blood samples, this protocol has been validated with fresh blood collected in EDTA collection tubes. Other anticoagulants may be used but are not guaranteed when using this kit.
For saliva samples, this protocol is validated for saliva collected only in Oragene DNA Saliva collection tubes.
The products previously known as Illumina DNA Prep with Exome 2.0 Plus Enrichment are now called Illumina DNA Prep with Exome 2.5 Enrichment (Cat. No. 20077595 and 20077596). The exome panel oligos have not changed, but the included indexes have been updated to Illumina DNA/RNA UD Indexes.
The new Illumina DNA Prep with Exome 2.5 Enrichment v2 kits (Cat. No. 20155541 and 20155543) also contain the same exome panel content with improved reagents that allow better coverage over GC-rich target areas.
Targeted custom enrichment panels, providing simple and efficient solutions for multiple targeted sequencing applications.
Advanced chemistry, optics, and informatics combine to deliver exceptional sequencing speed and data quality, outstanding throughput, and scalability.
Streamlined RNA-Seq solution for clear and comprehensive analysis across the coding and non-coding transcriptome with exceptional study flexibility.
A complete targeted resequencing solution for a wide range of applications, with fast integrated workflow and options for custom and fixed enrichment panels.
A fast, flexible, and scalable human whole-exome sequencing solution with support for low-input samples.
Enable CGP with a large pan-cancer panel covering all main variant classes plus key biomarkers (TMB, MSI, and HRD) from FFPE tissue.
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