To provide greater flexibility in ordering single components for libray preparation, we are moving to a modular configuration. Our existing TruSeq DNA PCR-Free Library Prep Kits (Cat. No. FC-121-3001, FC-121-3002, and FC-121-3003) will transition to TruSeq DNA PCR-Free (Cat. No. 20015962 for 24 samples and Cat. No. 20015963 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 for 96 indexes).

TruSeq DNA PCR-Free

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

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



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



Index Adapters

TruSeq DNA Single Indexes Set A (12 Indexes, 24 Samples)



TruSeq DNA Single Indexes Set B (12 Indexes, 24 Samples)



TruSeq DNA CD Indexes (96 Indexes, 96 Samples)



IDT for Illumina – TruSeq DNA UD Indexes (24 Indexes, 96 Samples)



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


Accessory Products

Illumina Free Adapter Blocking Reagent (12 reactions)



Illumina Free Adapter Blocking Reagent (48 reactions)



Product Highlights

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

Within our whole-genome sequencing workflows, 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.

Find a list of automation vendors with robotic systems that support the HT workflows


Method-Specific Workflow Example


Customer Stories

Feline Social Media Star Gets Sequenced

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 More
Single-Cell Transcriptome Sequencing, the Internet, and Memory

TGen researchers are combining crowd-sourcing and single-cell RNA sequencing on HiSeq Systems to adequately power genetic studies of memory.

Read More

Product Comparison

Comparison of TruSeq Library Preparation Kits
Product TruSeq DNA PCR-Free TruSeq DNA Nano
Product versions Low-Throughput (LT) and High-Throughput (HT) LT and HT
Description Comprehensive genomic coverage with reduced library bias and coverage gaps (compared to PCR-based options) Based upon widely adopted TruSeq library prep, with lower input and improved data quality (compared to prior TruSeq kit versions)
Automation options 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 1–2 μg 100–200 ng
Includes PCR No Yes
Assay time ~5 hours 5.5 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, and HiScanSQ systems

Product Literature

Comparison of TruSeq Library Preparation Kits

Technical Note | PDF < 1 MB

TruSeq DNA PCR-Free

Data Sheet | PDF 8 MB

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