Overview
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.
Key features
Provides rapid library preparation in as little as 90 min with only 15 min of hands-on time
Eliminates library quantification before sample pooling and sequencing
Delivers quick results in under 8 hr using Illumina sequencing systems
Achieves high, even coverage and accurate calls for PCR amplicons
Produces high-quality de novo assembly of small genomes
Allows scalability by processing and uniquely barcoding up to 384 samples in a single experiment
Applications
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.
Example workflow
Nextera XT DNA Library Prep Kit
MiSeq System
MiniSeq System
NextSeq 550 System
DRAGEN Germline
DRAGEN Metagenomics Pipeline
DRAGEN Somatic
Local Run Manager
Project recommendations
Instrument: Recommended number of samples
MiSeq System: Up to 384 samples per run
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
Related applications and methods
Microbial sequencing methods
NGS-based microbial sequencing methods include shotgun metagenomics, 16S rRNA sequencing, whole-genome and de novo sequencing, and transcriptomics.
Whole-genome sequencing (WGS)
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, or scRNA-Seq, you can study cellular differences often masked by bulk sampling. Explore high- and low-throughput single-cell sequencing methods.
High coverage, accurate calls
Nextera XT kit produces high and even coverage across a range of amplicon sizes with accurate variant calls as confirmed within the dbSNP database.
Even coverage across large amplicons
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.
High-quality de novo assembly of small genomes
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.
Best practices for Nextera library prep
This video reviews the key steps in the Nextera library preparation process and provides some tips and tricks on how to achieve optimal results.
Nextera XT: Sample preparation and usage
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. View less
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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. View less
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). View less
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). View less
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. View less
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. View less
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. View less
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. View less
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. View less
Nextera XT Index Kit v2 Set B (96 indexes, 384 samples)
FC-131-2002
Nextera XT Index Kit v2 Set C (96 indexes, 384 samples)
FC-131-2003
Nextera XT Index Kit v2 Set D (96 indexes, 384 samples)
FC-131-2004
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. View less
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. View less
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. View less
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How does Nextera tagmentation work?
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.
What is the difference between Nextera and TruSeq chemistries?
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.
What is the maximum number of samples that can be multiplexed per Nextera XT library?
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