To provide greater flexibility in ordering single components for library preparation, we are moving to a modular configuration. Our existing TruSeq Nano DNA Library Prep Kits (Cat. No. FC-121-4001, FC-121-4002, and FC-121-4003) will transition to TruSeq DNA Nano (Cat. No. 20015964 for 24 samples and Cat. No. 20015965 for 96 samples) for library preparation. Index adapters will be sold separately as TruSeq DNA Single Indexes (Cat. No. 20015960 for Set A and Cat. No. 20015961 for Set B) and TruSeq DNA CD (combinatorial dual) Indexes (Cat. No. 20015949).
TruSeq DNA Nano enables efficient interrogation of samples with limited available DNA. Based on mechanical fragmentation–ligation chemistry, this low-input method delivers high coverage quality and reduced bias for virtually any sequencing application.
TruSeq DNA Nano has been streamlined by replacing gel-based size selection with bead-based selection, enabling researchers to prepare high-quality libraries in less than a day. It is optimized for a variety of read lengths, from 2 × 101 bp to 2 × 151 bp.
TruSeq DNA Nano offers excellent results with as little as 100 ng input, eliminating the typical requirement for micrograms of DNA. This enables researchers to study samples with limited available DNA (eg, tumor samples) and helps preserve samples for use in future or alternate studies. In addition to accelerating the workflow, bead-based size selection avoids typical sample loss associated with gel-based selection.
TruSeq DNA Nano reduces the number and average size of typical PCR-induced gaps in coverage. The enhanced workflow reduces library bias and improves coverage uniformity across the genome. These kits also provide excellent coverage of traditionally challenging genomic content, including GC-rich regions, promoters, and repetitive regions. This enables researchers to access more information from each sequencing run. Truseq DNA Nano products are validated for high-quality genomic coverage for virtually any next-generation sequencing application.
View a list of automation vendors with robotic systems that support the 96-sample workflow
| 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 |
| HiSeq 2500 System | 2–10 samples per run (dual flow cell; based on 30× coverage of a human genome) | Rapid run, 350 bp insert: ≤ 2 × 100 bp Rapid run, 550 bp insert: ≤ 2 × 250 bp High output, 350 bp insert: ≤ 2 × 100 bp High output, 550 bp insert: ≤ 2 × 125 bp |
| TruSeq DNA Nano | TruSeq DNA PCR-Free | Nextera DNA Flex Library Prep Kit | |
|---|---|---|---|
| Assay Time | ~6 hours total assay time | 5 hours total assay time | ~3-4 hours (from DNA extraction to normalized library) |
| Automation Capability | Liquid Handling Robots | Liquid Handling Robots | Liquid Handling Robots |
| Description | A low-input research method that delivers high genome coverage quality and reduced bias. | A simple, all-inclusive PCR-free prep for whole-genome sequencing studies with the ability to sequence through challenging regions of the genome. | A fast, flexible workflow for a wide range of research applications and sample types, from human to microbial whole-genome sequencing and more. |
| Hands-On Time | ~4 hours | 4 hours | 1-1.5 hours |
| Input Quantity | 100 ng genomic DNA | 1 ug DNA | 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). |
| Mechanism of Action | Mechanical DNA fragmentation and adapter ligation. Workflow uses reduced-bias PCR and is gel-free. | Mechanical DNA fragmentation and adapter ligation. Workflow is PCR-free and gel-free. | Bead-linked transposome |
| Method | Genotyping by Sequencing , Shotgun Sequencing , Whole-Genome Sequencing | Genotyping by Sequencing , Shotgun Sequencing , Whole-Genome Sequencing | Amplicon Sequencing , De Novo 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 24 single, 96 combinatorial (CD) dual, 24 unique dual and 96 unique dual (UD) combinations (available soon) | Up to 384 unique dual (UD) combinations and 96 combinatorial dual (CD) combinations |
| Specialized Sample Types | Low-Input Samples, Not FFPE-Compatible | Not FFPE-Compatible | Blood, Not FFPE-Compatible, Saliva |
| Species Category | Human, Mammalian, Mouse, Other, Plant, Rat | Human, Mammalian, Mouse, Other, Plant, Rat | Any Species, Bacteria, Drosophila, Human, Mammalian, Mouse, Nematode, Plant, Rat, Virus, Yeast, Zebrafish |
| Species Details | Compatible with most large DNA genomes. | Compatible with most large DNA genomes. | Compatible with any species |
| Target Insert Size | 350 bp or 550 bp | 350 bp or 550 bp | ~500 bp |
Karolinska Institutet researchers use Illumina sequencers to identify novel HPV types associated with non-melanoma skin cancers.
Read MoreBaseSpace Sequence Hub SRA Submission App enables tracking of samples, from library prep to data analysis, helping the Biotechnology Resource Center Sequencing Core (BRC-Seq) at the University of British Columbia deliver high-quality sequencing data and additional value to its customers.
Read More| Specification | TruSeq DNA Nano | TruSeq DNA PCR-Free |
|---|---|---|
| Kit type | TruSeq DNA Nano LT & HT | TruSeq DNA PCR-Free LT & HT |
| Description | Based upon widely adopted TruSeq library prep, with lower input and improved data quality | Superior genomic coverage with radically reduced library bias and gaps |
| Workflow | LT kits: Manual workflow
HT kits: Automation-friendly; manual protocols also available |
LT kits: Manual workflow
HT kits: Automation-friendly; manual protocols also available |
| Input quantity | 100–200 ng
|
1–2 μg |
| Includes PCR | Yes | No |
| Assay time | 5.5 hours | ~5 hours |
| Hands-on time | 4 hours | ~4 hours |
| Target insert size | 350 bp or 550 bp | 350 bp or 550 bp |
| Gel-free | Yes | Yes |
| Number of samples supported | 24 (LT) or 96 (HT) samples | 24 (LT) or 96 (HT) samples |
| Size-selection beads | Included | Included |
| Applications | Whole-genome sequencing applications, including whole-genome resequencing, de novo assembly, and metagenomics studies | |
| Sample multiplexing | Up to 24 single, 96 combinatorial (CD) dual, 24 unique dual and 96 unique dual (UD) combinations | |
| Compatible Illumina sequencers | MiniSeq, MiSeq, NextSeq, HiSeq, NovaSeq, HiSeq X, HiScanSQ, and Genome Analyzer systems | |
Comparison of TruSeq Sample Preparation Kits
Technical Note | PDF< 1 MB
Mechanical DNA Fragmentation with the Q800R2 Sonicator
Technical Note | PDF4 MB
Data Sheet | PDF11 MB
TruSeq Nano DNA Library Prep Reference Guide Documentation
TruSeq HT Dual Index Quick Reference Card Documentation
TruSeq DNA Nano Reference Guide Documentation
TruSeq DNA Nano Checklist Documentation
TruSeq Nano DNA Library Prep User-Supplied Consumables and Equipment Documentation
TruSeq DNA Nano Consumables & Equipment Documentation
TruSeq Nano DNA Library Prep Checklist Documentation
Illumina Experiment Manager User Guide 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
Simple, all-inclusive whole-genome sequencing (WGS) library preparation that provides accurate and comprehensive coverage of complex genomes.
Prepare sequencing libraries for small genomes, PCR amplicons, plasmids, or cDNA in as little as 90 minutes, with a low DNA input requirement.
A fast, integrated workflow for a wide range of applications, from human whole-genome sequencing to amplicons, plasmids, and microbial species.
