Overview
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).
Get comprehensive coverage of the human exome with this focused panel offering high coverage for variants in reported public databases
Improve coverage of GC-rich targets with an updated workflow offering reduced GC dropout and increased coverage uniformity
Save time and effort with a rapid, easy to use whole-exome sequencing (WES) workflow, offering built-in yield normalization and a fast protocol combining fragmentation and indexing steps
Add or boost target coverage with spike-in panels with ease—choose mitochondrial genome coverage, or design a custom panel focused on your targets of interest
Customize your exome content
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.
View Illumina Custom Enrichment Panel v2
Whole-exome sequencing workflow from a single vendor
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.
What panels do I need?
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.
Applications
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.
Example workflow
Illumina DNA Prep with Exome 2.5 Enrichment v2
NextSeq 550 System
NextSeq 1000 & 2000 Systems
NovaSeq 6000 System
NovaSeq X Series
DRAGEN Enrichment
Emedgene software
Illumina Connected Insights - Somatic
Not available in all countries. Illumina Connected Insights supports user-defined tertiary analysis through API calls to third-party knowledge sources.
Project recommendations
Instrument: Recommended number of samples
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)
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.
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.
Related applications and methods
Whole-exome sequencing
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.
Cancer genomics research
Our sequencing and microarray technologies support a broad range of cancer genomics research applications, from DNA to RNA analysis, epigenetics, and more.
Figures
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.
Video resources
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). View less
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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). View less
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Is it possible to use < 50 ng of gDNA with Illumina DNA Prep with Exome 2.5 Enrichment v2?
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.
What blood or saliva collection tubes can be used with Illumina DNA Prep with Exome 2.5 Enrichment v2?
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.
What happened to Illumina DNA Prep with Exome 2.0 Plus Enrichment products?
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.
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