To provide greater flexibility in ordering single components for library 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...
What products do I need?
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)



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

Frequently Purchased Together



仪器 推荐的样本数 读长
NextSeq 550 System 每次运行1个样本(基于30倍的人类基因组覆盖度) 350 bp 插入片段: ≤ 2 × 100 bp
550 bp 插入片段: ≤ 2 × 150 bp
HiSeq 2500 System 每次运行2–10个样本(双流动槽;基于30倍的人类基因组覆盖度) 350 bp 插入片段: ≤ 2 × 100 bp (快速运行)
550 bp 插入片段: ≤ 2 × 250 bp (快速运行)
350 bp 插入片段: ≤ 2 × 100 bp (高通量)
550 bp 插入片段: ≤ 2 × 125 bp (高通量)
NovaSeq 6000 System 每次运行的样本(双流动槽):S1:8,S2:16,S4:48(基于30倍的人类基因组覆盖度) 350 bp 插入片段: ≤ 2 × 150 bp
550 bp 插入片段: ≤ 2 × 150 bp


TruSeq DNA PCR-Free Nextera DNA Flex Library Prep Kit TruSeq DNA Nano
变异种类 染色体异常, 结构变异, 生殖系变异, 体细胞变异, 杂合性缺失(LOH), 拷贝数变异(CNV), 单核苷酸多态性(SNP), 基因融合 基因融合, 单核苷酸多态性(SNP), 体细胞变异, 拷贝数变异(CNV), 杂合性缺失(LOH), 插入缺失, 染色体异常, 生殖系变异, 结构变异 插入缺失, 拷贝数变异(CNV), 杂合性缺失(LOH), 染色体异常, 基因融合, 单核苷酸多态性(SNP), 体细胞变异, 生殖系变异, 结构变异
技术 测序 测序 测序
方法 鸟枪法测序 , 通过测序进行基因分型 , 全基因组测序 鸟枪法测序 , 全基因组测序 , 扩增子测序 , 从头测序 鸟枪法测序 , 通过测序进行基因分型 , 全基因组测序
物种类别 小鼠, 大鼠, 哺乳动物, 人类, 植物, 其他 人类, 所有物种, 病毒, 酵母, 植物, 大鼠, 细菌, 哺乳动物, 小鼠 植物, 大鼠, 小鼠, 哺乳动物, 人类, 其他
系统兼容性 MiSeq , MiSeq, 研究模式下的MiSeqDx , MiniSeq , NextSeq 550 , HiSeq 2000, HiSeq X Ten , NextSeq 500 , HiSeq 2500 , HiSeq 1000, HiSeq 4000 , HiSeq 3000 , HiSeq X Five , HiSeq 1500 HiSeq X Five , MiniSeq , HiSeq 2500 , NextSeq 550 , MiSeq , MiSeq, HiSeq X Ten , 研究模式下的MiSeqDx , HiSeq 4000 , HiSeq 3000 MiniSeq , HiSeq X Ten , HiSeq 4000 , HiSeq 3000 , HiSeq X Five , HiSeq 1500, MiSeq , MiSeq, NextSeq 550 , NextSeq 500 , HiSeq 2000, 研究模式下的MiSeqDx , HiSeq 1000, HiSeq 2500
自动化功能 自动液体处理设备 自动液体处理设备 自动液体处理设备

Method-Specific Workflow Example


Customer Stories


马普研究院和宾夕法尼亚大学的研究人员共同资助了Lil Bub的基因组测序,采用NextSeq 500系统揭示了此种猫罕见疾病的遗传因素。




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



Related Products

TruSeq DNA Nano

Generate whole-genome sequencing libraries and efficiently interrogate samples with limited available DNA.

Nextera DNA Flex Library Prep Kit

A fast, integrated workflow for a wide range of applications, from human whole-genome sequencing to amplicons, plasmids, and microbial species.

Nextera Mate Pair Library Prep Kit

This library preparation kit offers a gel-free method for preparing up to 12 kb mate pair libraries with minimal DNA input requirements.