Order the NovaSeq X Series
Advanced chemistry, optics, and informatics combine to deliver exceptional speed and data quality, outstanding throughput and scalability.
Offers a 90-min workflow to prepare DNA libraries for small genomes, PCR amplicons, plasmids, and cDNA sequencing, with a low DNA input requirement.
Assay time
Hands-on time
Input quantity
We recommend ordering the Illumina DNA/RNA UD Indexes Sets A-D (20091654, 20091656, 20091658, 20091660). The IDT for Illumina DNA/RNA UD Indexes Sets A-D (20027213, 20027214,20042666, 20042667) are being discontinued; the last order date is January 30th, 2025.
The Nextera XT DNA Library Preparation Kit offers a fast and easy workflow for preparing, quantifying, and normalizing libraries for sequencing small genomes, PCR amplicons, plasmids, microbial genomes, concatenated amplicons, and double-stranded cDNA from a low amount of input DNA (1 ng), making this kit amenable for use with precious samples available in limited quantity.
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.
Assay time | ~5.5 hr from DNA extraction to normalized library. (Library prep time: ~90 minutes). |
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Automation capability | Liquid handling robots |
Automation details | Explore available automation methods |
Description | Prepare sequencing libraries for small genomes, PCR amplicons, and plasmids in 90 min, with a low DNA input requirement. |
Hands-on time | 15 min |
Input quantity | 1 ng DNA |
Instruments | MiSeq System, iSeq 100 System, NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, MiniSeq System, NextSeq 500 System |
Mechanism of action | Enzymatic fragmentation |
Method | Amplicon sequencing, 16s rRNA sequencing, De novo sequencing, Whole-genome sequencing, Shotgun sequencing |
Multiplexing | Up to 384 uniquely indexed samples may be pooled and sequenced together. |
Nucleic acid type | DNA |
Specialized sample types | Not FFPE-compatible, Low-input samples, Single cells |
Species category | Drosophila, Any species, Mammalian, Mouse, Yeast, Zebrafish, Fungal, Human, Rat, Plant, Virus, Nematode, Bacteria |
Species details | Compatible with any species |
Target insert size | 300 bp–1.5 kb |
Technology | Sequencing |
Variant class | Single nucleotide polymorphisms (SNPs), Structural variants |
Required products:
Illumina Purification Beads for purifying amplified libraries.
One of the following index kits:
Bead-based normalization:
Standard normalization:
The Nextera XT DNA Library Preparation Kit supports a low DNA input (1 ng) and offers high-quality sequencing-ready libraries for small genomes, PCR amplicons, and plasmids, in as little as 90 minutes.
Nextera XT DNA Library Prep Kit
Instrument | Recommended number of samples | Read length |
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MiSeq System | Up to 384 samples per run |
Up to 2 × 300 bp |
NextSeq 550 System | Up to 384 samples per run |
Up to 2 × 150 bp |
NGS-based microbial sequencing methods include shotgun metagenomics, 16S rRNA sequencing, whole-genome and de novo sequencing, and transcriptomics.
Whole-genome sequencing delivers a comprehensive view, ideal for discovery applications. Newer genome sequencers perform WGS more rapidly than ever.
Single-cell and ultra-low-input RNA-Seq
With single-cell RNA-Seq, you can study cellular differences often masked by bulk sampling. Explore high- and low-throughput single-cell analysis methods.
Nextera XT DNA Library Preparation Kit | Illumina DNA Prep | |
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Assay time | ~5.5 hr from DNA extraction to normalized library. (Library prep time: ~90 minutes). | ~3-4 hr (from DNA extraction to normalized library) |
Automation capability | Liquid handling robots | Liquid handling robots |
Automation details | Explore available automation methods | Explore available automation methods |
Description | Prepare sequencing libraries for small genomes, PCR amplicons, and plasmids in 90 min, with a low DNA input requirement. | A fast, flexible library prep workflow that accommodates an assortment of sample types and DNA input amounts, allowing access to a wide range of applications, from human to microbial whole-genome sequencing and more. |
Hands-on time | 15 min | 1-1.5 hr |
Input quantity | 1 ng DNA | 1-500 ng DNA for small genomes (e.g. microbial), or 100-500 ng DNA for large genomes (e.g. human). (For blood and saliva, see the reference guide). |
Instruments | MiSeq System, iSeq 100 System, NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, MiniSeq System, NextSeq 500 System | MiSeq System, iSeq 100 System, NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, MiSeqDx in Research Mode, MiniSeq System, NextSeq 550Dx in Research Mode, NovaSeq X System, NovaSeq 6000 System, NovaSeq X Plus System |
Mechanism of action | Enzymatic fragmentation | Bead-linked transposome |
Method | Amplicon sequencing, 16s rRNA sequencing, De novo sequencing, Whole-genome sequencing, Shotgun sequencing | Amplicon sequencing, De novo sequencing, Whole-genome sequencing, Shotgun 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 |
Nucleic acid type | DNA | DNA |
Specialized sample types | Not FFPE-compatible, Low-input samples, Single cells | Blood, Not FFPE-compatible, Saliva |
Species category | Drosophila, Any species, Mammalian, Mouse, Yeast, Zebrafish, Fungal, Human, Rat, Plant, Virus, Nematode, Bacteria | Drosophila, Any species, Mammalian, Mouse, Yeast, Zebrafish, Human, Rat, Plant, Virus, Nematode, Bacteria |
Species details | Compatible with any species | Compatible with any species |
Target insert size | 300 bp–1.5 kb | ~350 bp |
Technology | Sequencing | Sequencing |
Variant class | Single nucleotide polymorphisms (SNPs), Structural variants | Single nucleotide polymorphisms (SNPs), Gene fusions, Loss of heterozygosity (LOH), Somatic variants, Chromosomal abnormalities, Germline variants, Structural variants, Insertions-deletions (indels), Copy number variants (CNVs) |
Library Prep and Array Kit Selector
Find the right sequencing library preparation kit or microarray for your needs. Filter by method, species, and more. Compare, share, and order kits.
DesignStudio Custom Assay Design Tool
DesignStudio is a web-based assay design tool to help scientists design and order custom sequencing probes or create custom genotyping array assays.
Nextera XT kit produces high and even coverage across a range of amplicon sizes with accurate variant calls as confirmed within the dbSNP database.
Nextera XT kit shows (A) high sequencing coverage (> 1000×) and (B) accurate variant calling along positions of a large amplicon (5.1 kb) in a highly variable non-coding region of the human genome.
Nextera XT kit produces high-quality de novo assembly of Escherichia coli genome with excellent N50 scores and coverage from 1 ng of genomic DNA.
This video reviews the key steps in the Nextera library preparation process and provides some tips and tricks on how to achieve optimal results.
This webinar on the Nextera XT sample preparation process will cover the protocol, best practices for critical steps, and recommended applications.
Nextera XT DNA Library Preparation Kit (24 samples)
FC-131-1024
Includes reagents for preparing 24 libraries. Purchase index adapters and purification beads separately.
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Nextera XT DNA Library Preparation Kit (96 samples)
FC-131-1096
Includes reagents for preparing 96 libraries. Purchase index adapters and purification beads separately.
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TG Nextera® XT DNA Sample Preparation Kit (96 Samples)
TG-131-1096
Each Nextera® XT DNA Sample Preparation kit provides reagents sufficient for preparing up to 96 samples.
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TG Nextera® XT Index Kit v2 Set A (96 Indices, 384 Samples)
TG-131-2001
Version 2 of 96 indexes for Nextera XT sample prep (Set A).
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TG Nextera® XT Index Kit v2 Set B (96 Indices, 384 Samples)
TG-131-2002
Version 2 of 96 indexes for Nextera XT sample prep (Set B).
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Illumina® DNA/RNA UD Indexes Set A, Tagmentation (96 Indexes, 96 Samples)
20091654
Illumina® DNA/RNA UD Indexes Set A, Tagmentation (96 Indexes, 96 Samples) Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.
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Illumina® DNA/RNA UD Indexes Set B, Tagmentation (96 Indexes, 96 Samples)
20091656
Illumina® DNA/RNA UD Indexes Set B, Tagmentation (96 Indexes, 96 Samples) Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.
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Illumina® DNA/RNA UD Indexes Set C, Tagmentation (96 Indexes, 96 Samples)
20091658
Illumina® DNA/RNA UD Indexes Set C, Tagmentation (96 Indexes, 96 Samples) Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.
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Illumina® DNA/RNA UD Indexes Set D, Tagmentation (96 Indexes, 96 Samples)
20091660
Illumina® DNA/RNA UD Indexes Set D, Tagmentation (96 Indexes, 96 Samples) Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.
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IDT® for Illumina® DNA/RNA UD Indexes Set A, Tagmentation (96 Indexes, 96 Samples)
20027213
Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.
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IDT® for Illumina® DNA/RNA UD Indexes Set B, Tagmentation (96 Indexes, 96 Samples)
20027214
Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.
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IDT® for Illumina® DNA/RNA UD Indexes Set C, Tagmentation (96 Indexes, 96 Samples)
20042666
Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.
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IDT® for Illumina® DNA/RNA UD Indexes Set D, Tagmentation (96 Indexes, 96 Samples)
20042667
Includes 96, 10 bp indexes sufficient for labeling 96 samples. Purchase library prep and enrichment reagents and probe panels separately.
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Nextera XT Index Kit v2 Set A (96 indexes, 384 samples)
FC-131-2001
Includes 96, 8 bp indexes sufficient for labeling 384 samples. Purchase library prep kit and purification beads separately.
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Nextera XT Index Kit v2 Set B (96 indexes, 384 samples)
FC-131-2002
Includes 96, 8 bp indexes sufficient for labeling 384 samples. Purchase library prep kit and purification beads separately.
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Nextera XT Index Kit v2 Set C (96 indexes, 384 samples)
FC-131-2003
Includes 96, 8 bp indexes sufficient for labeling 384 samples. Purchase library prep kit and purification beads separately.
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Nextera XT Index Kit v2 Set D (96 indexes, 384 samples)
FC-131-2004
Includes 96, 8 bp indexes sufficient for labeling 384 samples. Purchase library prep kit and purification beads separately.
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Nextera XT Index Kit (24 indexes, 96 samples)
FC-131-1001
Includes 24, 8 bp indexes sufficient for labeling 96 samples. Purchase library prep and purification beads separately.
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Illumina Purification Bead, 100mL
20060057
Includes one, 100-ml container of beads for library clean-up and size selection. Purchase library prep and enrichment reagents, probe panels, and indexes separately.
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Illumina Purification Bead, 400mL
20060058
Includes four, 100-ml container of beads for library clean-up and size selection. Purchase library prep and enrichment reagents, probe panels, and indexes separately.
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Nextera tagmentation uses in vitro transposition to prepare sequence-ready libraries from genomic DNA for all Illumina sequencing platforms. The tagmentation technology simultaneously fragments and tags DNA with sequencing adapters in a single-tube enzymatic reaction.
Nextera chemistry uses transposon-based DNA fragmentation whereby a transposon cleaves and tags DNA with adapters, while TruSeq chemistry follows an alternate enrichment protocol that uses mechanical shearing to fragment the DNA.
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