With Nextera technology, DNA is simultaneously fragmented and tagged with sequencing adapters in a single-tube enzymatic reaction. Nextera XT supports ultra-low DNA input of only 1 ng. It supports a wide range of input samples, including small genomes, PCR amplicons greater than 300 bp, plasmids, microbial genomes, concatenated amplicons, and double-stranded cDNA.
Multiplexing of up to 384 samples per Nextera XT library is available for projects requiring greater throughput. Plus, the kit includes an innovative bead-based sample normalization that eliminates the need for library quantification prior to pooling and sequencing. Libraries prepared with Nextera XT kits are compatible with all Illumina sequencers.
The IDT for Illumina-Nextera DNA UD Indexes Sets A, B, C, and D Indexes offer up to 384 unique dual indexing, which allows accurate assignment of reads and efficient use of the flow cell. These unique dual index codes use 10 bp codes. This change in base pair index codes requires adjustments to the sequencing run setup.
Find robotic systems compatible with this kit
This product is also available as an Illumina Advantage (TG) product. Illumina Advantage large-scale sequencing products feature lot-specific shipments and testing, extended shelf life, and advanced change notifications for greater laboratory efficiency.
For high-throughput research, bead-based normalization can save time and resources by providing an accelerated path from DNA to data. This technical note provides guidelines and data comparing the two normalization methods to help you decide which option to use.
Read Tech Note
| Instrument | Recommended Number of Samples | Read Length |
|---|---|---|
| MiSeq System | Up to 384 samples per run | Up to 2 x 300 bp |
| NextSeq 550 System | Up to 384 samples per run | Up to 2 x 150 bp |
| Nextera XT DNA Library Preparation Kit | Illumina DNA Prep | |
|---|---|---|
| Assay Time | ~5.5 hours from DNA extraction to normalized library. (Library prep time: ~90 minutes). | ~3-4 hours (from DNA extraction to normalized library) |
| Description | Fast library prep optimized for research on small genomes, PCR amplicons, and plasmids. | A fast, flexible workflow for a wide range of research applications and sample types, from human to microbial whole-genome sequencing and more. |
| Hands-On Time | 15 minutes | 1-1.5 hours |
| Input Quantity | 1 ng DNA | Small genomes (e.g. microbial): 1-500 ng DNA. Large genomes (e.g. human): 100-500 ng DNA. (For blood and saliva, see the reference guide). |
| Mechanism of Action | Enzymatic fragmentation | Bead-linked transposome |
| Method | 16s rRNA Sequencing, Amplicon Sequencing , De Novo Sequencing , Shotgun Sequencing , Whole-Genome Sequencing | Amplicon Sequencing , De Novo Sequencing , Shotgun Sequencing , Whole-Genome Sequencing |
| Multiplexing | Up to 384 uniquely indexed samples may be pooled and sequenced together. | Up to 384 unique dual (UD) combinations and 96 combinatorial dual (CD) combinations |
| Specialized Sample Types | Low-Input Samples, Not FFPE-Compatible, Single Cells | Blood, Not FFPE-Validated, Saliva |
| Species Details | Compatible with any species | Compatible with any species |
| Target Insert Size | 300 bp–1.5 kb | ~350bp |
Single-cell mRNA-Seq with the MiSeq and HiSeq Systems is enabling Allen Institute researchers to classify V1 neural cells.
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The combination of Nextera XT and rapid sequencing with the MiSeq System provides a comprehensive DNA to data workflow in only 8 hours.
The Nextera XT Library Preparation Kit eliminates the need for library quantification before sample pooling and sequencing. Libraries of equivalent concentrations are created by employing bead-based sample normalization, as simple as pipetting 5 μl of each library to be sequenced.
Nextera Library Validation and Cluster Density Optimization
Technical Note | PDF< 1 MB
Technical Note | PDF< 1 MB
Nextera XT Library Prep: Tips and Troubleshooting
Technical Note | PDF< 1 MB
Sequencing the Rapidly Evolving Influenza A Virus on the MiSeq System
Application Note | PDF< 1 MB
Nextera XT DNA Library Preparation Kit
Data Sheet | PDF< 1 MB
Nextera XT DNA Library Prep Kit Consumables & Equipment List Documentation
Nextera XT DNA Library Prep Kit Checklist Documentation
Nextera XT DNA Library Prep Kit Documentation
Illumina Experiment Manager User Guide Documentation
Index Adapters Pooling Guide Documentation
Nextera XT DNA Library Prep Kit Reference Guide Documentation
Custom Protocol Selector
Generates customized, end-to-end instructions
A fast, integrated workflow for a wide range of applications, from human whole-genome sequencing to amplicons, plasmids, and microbial species.
Optimized chemistry to increase cluster density and read length, and improve sequencing quality scores, compared to earlier MiSeq reagent kit versions.
iSeq 100 Reagents feature a ready-to-use cartridge and one-channel SBS technology for simple, low-cost sequencing.
