Simple, all-inclusive whole-genome sequencing (WGS) library preparation that provides accurate and comprehensive coverage of complex genomes.
Assay time
Hands-on time
Input quantity
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.
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:
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.
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.
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.
Prepare libraries for Illumina sequencing in less than a day. Illumina Qualified Methods are available on a range of automation platforms through our partners.
| Assay time | 5 hr total assay time |
|---|---|
| Automation capability | Liquid Handling Robots |
| Automation details | Explore available automation methods |
| Description | A simple, all-inclusive PCR-free prep for whole-genome sequencing studies with the ability to sequence through challenging regions of the genome. |
| Hands-on time | 4 hr |
| Input quantity | 1 ug 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 |
| Mechanism of action | Mechanical DNA fragmentation and adapter ligation. Workflow is PCR-free and gel-free. |
| Method | Shotgun Sequencing, Whole-Genome Sequencing, Genotyping by Sequencing |
| Multiplexing | Up to 24 single, 96 combinatorial (CD) dual, 24 unique dual and 96 unique dual (UD) combinations |
| Nucleic acid type | DNA |
| Specialized sample types | Not FFPE-Compatible |
| Species category | Other, Mammalian, Mouse, Human, Rat, Plant |
| Species details | Compatible with most large DNA genomes. |
| Target insert size | 350 bp or 550 bp |
| Technology | 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) |
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.
| 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 |
| 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 |
Large whole-genome sequencing informs disease research and population genomics studies and reveals disease-associated alleles.
Microbial Whole-Genome Sequencing
Use microbial whole-genome sequencing to map genomes of novel organisms, finish genomes of known organisms, or compare genomes across multiple samples.
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.
| 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 | |
| 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, 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 | Shotgun Sequencing, Whole-Genome Sequencing, Genotyping by Sequencing | Shotgun Sequencing, Whole-Genome Sequencing, De Novo Sequencing, Amplicon Sequencing | Shotgun Sequencing, Whole-Genome Sequencing, Genotyping by 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 | |
| Specialized sample types | Not FFPE-Compatible | Not FFPE-Compatible | Blood, Not FFPE-Compatible, Saliva | Not FFPE-Compatible, Low-Input Samples |
| Species category | Other, Mammalian, Mouse, Human, Rat, Plant | 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 | |
| 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) | 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) |
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.
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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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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