Nextera DNA Exome offers two configuration options. The product previously known as the TruSeq Rapid Exome Library Prep Kit (Cat. No. FC-144-1000, FC-144-1001, FC-144-1002, FC-144-1003, and FC-144-1004) is now called the Nextera DNA Exome Kit and is available as a pre-assembled kit. Kits include Illumina library preparation reagents, index adapters, enrichment reagents, and an exome panel. Alternatively, to combine Illumina Nextera DNA Exome library preparation with IDT xGen exome enrichment, major components (such as library preparation and index adapters) can be purchased separately. A full list of components available for individual purchase is shown on the “Choose Your Components” tab below. IDT xGen exome enrichment reagents may be purchased from IDT.
The Nextera Exome Kit, previously known as the TruSeq Rapid Exome Library Prep Kit, delivers a simple, efficient method for high-confidence calling of exonic variants. This fast library preparation and exome enrichment workflow delivers libraries in less than 2 days with no need for additional equipment, giving labs the flexibility to plan projects according to sample volume. Benefits include:
The Nextera Exome Kit delivers an average of >75% of on-target sequencing reads. This high on-target percentage requires fewer sequencing cycles to reach desired coverage levels but still achieves uniform coverage for high-confidence results.
To learn more about calculating coverage estimates, see the sequencing coverage calculator.
Illumina now offers modular product ordering to enable flexibility in your workflows.
Find a list of automation vendors with robotic systems compatible with this solution
Illumina has partnered with IDT to co-market a combined exome enrichment solution using the xGen® Exome Research Panel, which consists of 429,826 individually synthesized and quality-controlled xGen Lockdown® Probes. The panel spans a 39 Mb target region (19,396 genes) of the human genome, and covers 51 Mb of end-to-end tiled space.
xGen Blocking Oligos (Cat. No.1075474, 1075475, or 1075476 for TruSeq libraries; contact IDT for Nextera libraries) and xGen Lockdown Reagents (Cat. No. 1072280 or 1072281) are also required.
| Instrument | Recommended Number of Samples | Read Length |
|---|---|---|
| NextSeq 550 System | Samples per run: mid output: 3, high output: 12 (based on 50 million reads, 50× coverage, 4 Gb of data) | 2 × 75 bp (max recommended) |
| HiSeq 2500 System | Samples per run (dual flow cell): rapid run: 24, high output: 156 (based on 50 million reads, 50× coverage, 4 Gb of data) | 2 × 75 bp (max recommended) |
| NovaSeq 6000 System | Samples per run (dual flow cell): S1: 96, S2: 96, S4: 192 (based on 50 million reads, 50× coverage, 4 Gb of data) | ≤ 2 × 100 bp |
| Nextera DNA Exome | TruSeq DNA Exome | AmpliSeq for Illumina Exome Panel | Nextera Flex for Enrichment | |
|---|---|---|---|---|
| Assay Time | < 2.5 days | 2.5 days | < 7.5 hours (library prep only; doesn't include library quantification, normalization, or pooling time) | ~6.5 hours |
| Content Specifications | Captures coding exons. Covers 45 Mb of exonic content (≥98% of RefSeq, CCDS, and Ensembl coding content). | Captures coding exons. Covers 45 Mb of exonic content (≥98% of RefSeq, CCDS, and Ensembl coding content). | > 97% of CCDS | Exome: 45 Mb exonic content (≥98% of RefSeq, CCDS, and Ensembl coding content). Custom: 0.5 - 15 Mb genomic content of interest. Fixed panels: Content varies by panel. |
| Description | A cost-effective exome research workflow focused on ≥98% of RefSeq, CCDS, and Ensembl coding content. | A cost-effective exome research workflow focused on ≥98% of RefSeq, CCDS, and Ensembl coding content. | Enables researchers to prepare 8 exome libraries in <7.5 hours with <1.5 hours hands-on time. | A fast, flexible research solution that enables whole-exome, custom, and fixed panel targeted sequencing. |
| Hands-On Time | 3 hours | 6 hours | < 1.5 hours | ~2 hours |
| Input Quantity | 50 ng genomic DNA | 100 ng genomic DNA | 50 ng minimum | 10-1000 ng high-quality genomic DNA or 50-1000 ng FFPE DNA. (For blood and saliva, see the reference guide). |
| Mechanism of Action | Transposase-based fragmentation and exome enrichment with biotinylated capture probes | Mechanical fragmentation (Covaris) and exome enrichment with biotinylated capture probes | Multiplex PCR | Enrichment-compatible, bead-linked transposome |
| Method | Exome Sequencing | Exome Sequencing | Amplicon Sequencing , Exome Sequencing | Custom Sequencing, Exome Sequencing , Target Enrichment , Target Enrichment, Targeted DNA Sequencing |
| Multiplexing | Up to 12-plex enrichment | Up to 12-plex enrichment | 96 dual index combinations | Up to 384 available unique dual indexes. Pre-enrichment pooling of up to 12-plex is tested. |
| On-Target Reads | >75% | >80% | >80% | ≥85% (for exome panel) |
| Specialized Sample Types | Low-Input Samples, Not FFPE-Compatible | FFPE Tissue | Not FFPE-Compatible | Blood, FFPE Tissue, Saliva |
| Species Category | Human | Human | Human | Human, Other |
| Uniformity | >75% (% coverage at 20x for 4 Gb) | >85% (% coverage at 20x for 4 Gb) | >90% (% coverage at 20x for 4 Gb) | ≥90% (% coverage at 20x for 4 Gb; for exome panel) |
Y-axis: Fluorescence Units (FU), X-axis: Base Pairs (bp)
Enhanced transposome chemistry is highly tolerant of varying DNA input amounts, reducing bias and providing consistent results.
| Catalog Number | Total Samples | Number of Enrichment Reactions | Index Adapters |
|---|---|---|---|
| 20020616 | 24 | 8 | 24 combinatorial dual indexes |
| 20020617 | 96 | 8 | 96 combinatorial dual indexes |
Illumina Exomes Comparative Table
Product Information Sheet | PDF< 1 MB
Data Sheet | PDF | 5 versions
TruSeq Rapid Exome Library Prep Reference Guide Documentation
TruSeq Rapid Exome Library Prep Checklist Documentation
Nextera DNA Exome Documentation
Nextera DNA Exome Reference Guide Documentation
Nextera DNA Exome Support - Documentation & Literature
Sequencing Library qPCR Quantification Guide Documentation
Nextera DNA Exome Checklist Documentation
TruSeq Rapid Exome Library Prep Consumables and Equipment List Documentation
Nextera DNA Exome Consumables & Equipment Documentation
Custom Protocol Selector
Generates customized, end-to-end instructions
Nextera DNA Exome (formerly known as TruSeq Rapid Exome) is the recommended replacement product for the Nextera Rapid Capture Exome and Expanded Exome Kits, which have been discontinued. Illumina remains committed to providing you with high-quality support and service.
This kit provides a low-cost exome sequencing solution that delivers exceptional target coverage over a broad range of read depths.
A fast, integrated workflow for a wide range of applications providing targeted resequencing for custom panels, fixed panels, and whole-exome enrichment.
Comprehensive sequencing research panels targeting disease-associated regions of the exome with high analytical sensitivity and specificity.
Targeted research panel for sequencing the protein-coding regions of the human genome.
