TruSeq DNA PCR-Free

Simple, all-inclusive whole-genome sequencing (WGS) library preparation that provides accurate and comprehensive coverage of complex genomes.

5 hr total assay time

Assay time

4 hr

Hands-on time

1 ug DNA

Input quantity

See full details in the specifications table

Illumina DNA PCR-Free Prep is now available. This library prep kit achieves commensurate performance with TruSeq DNA PCR-Free, while reducing turnaround time and hands-on time.

Overview

TruSeq DNA PCR-Free provides simple, all-inclusive library preparation for whole-genome sequencing applications. Researchers can sequence a wide variety of organisms, from small genomes such as bacteria to human whole genomes. The workflows offer:

  • Shortened gel-free workflows that remove the need for PCR

  • Ability to sequence challenging regions

  • Excellent coverage quality for deep insight into the genome

Sequence challenging regions

TruSeq DNA PCR-Free offers superior coverage of areas that are traditionally difficult to sequence, such as GC-rich regions, promoters, and repetitive content.

The workflows are tunable to a variety of read lengths and are supported on all Illumina sequencing instruments. This permits the researcher to tailor each run to the needs of the experiment.

Gain deep insight into the genome

PCR-free means reduced library bias and gaps. The result is high data quality and optimal variant detection across the genome. Excellent genome coverage quality means your results have few gaps and good coverage of GC-rich regions.

Save time with a PCR-free protocol

Removing PCR saves time and removes genomic coverage bias associated with PCR steps. Bead-based size selection shortens the workflow. In tandem with Illumina sequencing systems, TruSeq DNA PCR-Free provides a range of enhancements to a widely adopted library preparation workflow.

Access flexible throughput options

  • TruSeq DNA PCR-Free with Single Indexes supports 24-sample manual processing for low-throughput (LT) studies.
  • TruSeq DNA PCR-Free with 96 CD Indexes supports 96-sample processing for high-throughput (HT) studies, and can be automated on liquid-handling robots (or processed manually).
  • IDT for Illumina Unique Dual (UD) Indexes (24 or 96) offer increased plexity that enables accurate assignment of reads and efficient use of the flow cell.

Fast, Automation-friendly Workflow

Prepare libraries for Illumina sequencing in less than a day. Illumina Qualified Methods are available on a range of automation platforms through our partners.


Specifications


Required products

Illumina now offers modular product ordering to enable flexibility in your workflows. You may order any of the following library prep products with any of the following indexes. 

/ Results

Applications

Example workflow


Project recommendations

Instrument Recommended number of samples Read length
NextSeq 550 System

1 sample per run (based on 30× coverage of a human genome) 

350 bp insert: ≤ 2 × 100 bp
550 bp insert: ≤ 2 × 150 bp

NovaSeq 6000 System

Samples per run (dual flow cell): S1: 8, S2: 16, S4: 48 (based on 30× coverage of a human genome) 

350 bp insert: ≤ 2 × 150 bp
550 bp insert: ≤ 2 × 150 bp


Related applications and methods

How others use this product

Researchers from the Max Planck Institute and the University of Pennsylvania crowdfunded the genome sequencing of Lil Bub, using the NextSeq 500 System to uncover the genetic causes of the cat’s rare diseases.

Compare

TruSeq DNA PCR-Free Illumina DNA PCR-Free Prep Illumina DNA Prep TruSeq DNA Nano
Assay time 5 hr total assay time ~1.5 hr ~3-4 hr (from DNA extraction to normalized library)

~6 hr total assay time

Automation capability Liquid handling robots Liquid handling robots Liquid handling robots Liquid handling robots
Automation details Explore available automation methods Explore available automation methods Explore available automation methods Explore available automation methods
Hands-on time 4 hr ~45 min 1-1.5 hr ~4 hr
Input quantity 1 ug DNA 25 ng to 300 ng 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). 100 ng genomic DNA
Instruments MiSeq System, NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, MiSeqDx in Research Mode, MiniSeq System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System MiSeq System, NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, MiSeqDx in Research Mode, NextSeq 550Dx in Research Mode, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus 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 MiSeq System, NextSeq 550 System, NextSeq 2000 System, MiSeqDx in Research Mode, MiniSeq System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System
Mechanism of action Mechanical DNA fragmentation and adapter ligation. Workflow is PCR-free and gel-free. Bead-linked transposome Bead-linked transposome Mechanical DNA fragmentation and adapter ligation. Workflow uses reduced-bias PCR and is gel-free.
Method Genotyping by sequencing, Whole-genome sequencing, Shotgun sequencing Whole-genome sequencing, De novo sequencing Amplicon sequencing, Whole-genome sequencing, De novo sequencing, Shotgun sequencing Genotyping by sequencing, Whole-genome sequencing, Shotgun sequencing
Multiplexing Up to 24 single, 96 combinatorial (CD) dual, 24 unique dual and 96 unique dual (UD) combinations Up to 384-plex Up to 384 unique dual (UD) combinations and 96 combinatorial dual (CD) combinations Up to 24 single, 96 combinatorial (CD) dual, 24 unique dual and 96 unique dual (UD) combinations
Nucleic acid type DNA DNA DNA DNA
Specialized sample types Not FFPE-Compatible Blood, Not FFPE-Compatible, Low-Input Samples, Saliva, Not FFPE-Compatible Blood, Not FFPE-Compatible, Saliva Not FFPE-Compatible, Low-Input Samples
Species category Other, Mammalian, Mouse, Human, Rat, Plant Any species, Human Drosophila, Any species, Mammalian, Mouse, Yeast, Zebrafish, Human, Rat, Plant, Virus, Nematode, Bacteria Other, Mammalian, Mouse, Human, Rat, Plant
Species details Compatible with most large DNA genomes. Compatible with any species Compatible with most large DNA genomes.
Target insert size 350 bp or 550 bp 450 bp +/- 75 bp ~350 bp 350 bp or 550 bp
Technology Sequencing Sequencing Sequencing Sequencing
Variant class 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) Short tandem repeats (STRs), Single nucleotide polymorphisms (SNPs), Somatic variants, Chromosomal abnormalities, Germline variants, Structural variants, Insertions-deletions (indels), Copy number variants (CNVs) 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) 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)

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Library prep (2)

TruSeq DNA PCR-Free Low Throughput Library Prep Kit (24 samples)

20015962

Includes reagents and Sample Purification Beads for preparing 24 libraries. Purchase index adapters separately.

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TruSeq DNA PCR-Free High Throughput Library Prep Kit (96 samples)

20015963

Includes reagents and Sample Purification Beads for preparing 24 libraries. Purchase index adapters separately.

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Index adapters (1)

IDT for Illumina – TruSeq DNA UD Indexes v2 (96 Indexes, 96 Samples)

20040870

Includes 96, 8 bp indexes sufficient for labeling 96 samples and is compatible with TruSeq Nano and TruSeq PCR-Free Library Prep Kits. Version 2 contains improvements for 8 of the 192 index oligos in plate 20022370, with no changes to the remaining 184. Purchase library preps and probe panels separately.

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Product

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Unit Price

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