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.
| 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 |
| TruSeq DNA PCR-Free | Illumina DNA PCR-Free Prep | Illumina DNA Prep | TruSeq DNA Nano | |
|---|---|---|---|---|
| Assay Time | 5 hours total assay time | ~1.5 hours | ~3-4 hours (from DNA extraction to normalized library) | ~6 hours total assay time |
| Automation Capability | Liquid Handling Robots | Liquid Handling Robots | Liquid Handling Robots | Liquid Handling Robots |
| Hands-On Time | 4 hours | ~45 minutes | 1-1.5 hours | ~4 hours |
| Input Quantity | 1 ug DNA | 25 ng to 300 ng | 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). | 100 ng genomic DNA |
| 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, Shotgun Sequencing, Whole-Genome Sequencing | De Novo Sequencing, Whole-Genome Sequencing | Amplicon Sequencing, De Novo Sequencing, Shotgun Sequencing, Whole-Genome Sequencing | Genotyping by Sequencing, Shotgun Sequencing, Whole-Genome Sequencing |
| Multiplexing | Up to 24 single, 96 combinatorial (CD) dual, 24 unique dual and 96 unique dual (UD) combinations (available soon) | 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 (available soon) |
| Specialized Sample Types | Not FFPE-Compatible | Blood, Low-Input Samples, Not FFPE-Compatible, Saliva | Blood, Not FFPE-Validated, Saliva | Low-Input Samples, Not FFPE-Compatible |
| Species Category | Human, Mammalian, Mouse, Other, Plant, Rat | Any Species, Human | Any Species, Bacteria, Drosophila, Human, Mammalian, Mouse, Nematode, Plant, Rat, Virus, Yeast, Zebrafish | Human, Mammalian, Mouse, Other, Plant, Rat |
| 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 | ~350bp | 350 bp or 550 bp |
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.
Read MoreData Sheet | PDF | 5 versions
Mechanical DNA Fragmentation with the Q800R2 Sonicator
Technical Note | PDF 4 MB
Comparison of TruSeq Sample Preparation Kits
Technical Note | PDF < 1 MB
TruSeq HT Dual Index Quick Reference Card Documentation
Illumina Experiment Manager User Guide Documentation
TruSeq DNA PCR-Free Reference Guide Documentation
TruSeq DNA PCR-Free Checklist Documentation
TruSeq DNA PCR-Free Library Prep User-Supplied Consumables and Equipment Documentation
TruSeq DNA PCR-Free Library Prep Checklist Documentation
TruSeq DNA PCR-Free Consumables & Equipment Documentation
Custom Protocol Selector
Generates customized, end-to-end instructions
Determine the best library prep kit or array for your needs based on your starting material and method of interest.
Access Tool
Generate whole-genome sequencing libraries and efficiently interrogate samples with limited available DNA.
The NovaSeq Xp workflow enables labs to load libraries directly into each lane of a NovaSeq 6000 System flow cell, without additional instrumentation.
A fast, integrated workflow for a wide range of applications, from human whole-genome sequencing to amplicons, plasmids, and microbial species.
A high-performing, fast, and integrated workflow for sensitive applications such as human whole-genome sequencing.
