QIAseq 1-Step Amplicon Library Kit

For fast and efficient preparation of DNA libraries for use in NGS applications
  • One-tube, benchtop library prep from PCR products in just 30 minutes
  • Maximal convenience with automatable, 1-reaction, room-temperature setup
  • High-quality, artifact-free libraries, ready for use on any Illumina NGS platform
  • Compatible with any gene panel or PCR product, only needs 1 ng input DNA
  • Single-use adapter plates minimize risk of contamination or handling errors
Save time with targeted resequencing and amplicon sequencing applications by taking advantage of the fastest library prep solution on the market – the QIAseq 1-Step Amplicon Library Kit. By combining end-repair and ligation, this new kit offers a fast and efficient 30-minute procedure allowing you to prepare high-quality, artifact-free libraries from any QIAGEN GeneRead Targeted DNA Panel and Ampli-Seq, Tru-Seq or PCR amplicons. The resulting NGS library can be used on any Illumina NGS instrument.
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QIAseq 1-Step Amplicon Library Kit (12)

For 12 reactions: 1-Step Amplicon Enzyme Mix, 4x 1-Step Amplicon Buffer, Primer Mix Illumina Library Amp, HiFi PCR Master Mix, RNase-Free Water. Adapters sold separately.

180412
313,00 €
QIAseq 1-Step Amplicon Library Kit (96)

For 96 reactions: 1-Step Amplicon Enzyme Mix, 4x 1-Step Amplicon Buffer, Adapter Plate 96-plex Illumina, Primer Mix Illumina Library Amp, HiFi PCR Master Mix, RNase-Free Water

180415
2.811,00 €
The QIAseq 1-Step Amplicon Library Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Optimized One-Step Library Construction.|Overview of a Complete Targeted Resequencing NGS Workflow Including the QIAseq 1-Step Amplicon Library Kit.|Typical Results.|
The QIAseq 1-Step Amplicon Library Kit offers substantial time savings over standard targeted resequencing library preparations. Purified amplicons from multiplexed PCR or gene panels are converted to sequencing libraries by employing a 30-minute, one-tube library construction step. The libraries are purified with an easy and automatable size selection protocol, and the entire procedure can be performed at room temperature. For low-input applications, libraries can be amplified using the included HiFi PCR Master Mix.|After DNA extraction and purification with an appropriate kit, target enrichment is performed with the QIAGEN Multiplex PCR Kit, GeneRead DNAseq Targeted Panels V2 or other gene panels and PCR products. NGS library construction is performed with the QIAseq 1-Step Amplicon Library Kit and data is analyzed with the CLC Genomics or Biomedical Workbench.|

In this experiment, the QIAGEN GeneRead DNAseq V2 Human Comprehensive Cancer Panel (cat. no. 181901) was used to generate PCR products [A] from a reference DNA sample. [B] A library was prepared from 25 ng of purified PCR product using the QIAseq 1-Step Amplicon Library Kit. Only 4 amplification cycles after library preparation were applied. Note that libraries display an appropriate size-shift and are free from adapter-adapter products (~125 bp).|

Performance
Significant time savings – from amplicon to high-quality, NGS-ready library in just 30 minutes
Traditional NGS library prep can be laborious, taking anything from 2 to 3 hours to accomplish. The QIAseq 1-Step Amplicon Library Kit offers a faster, more efficient alternative, allowing reliable NGS library prep from pools of PCR fragments from gene panels or multiplexed PCR. Utilizing a novel, combined end-repair/ligation reaction, the kit streamlines the entire NGS library preparation process to just 30 minutes (see figures Optimized One-step Library Construction and Overview of a Complete Targeted Resequencing NGS Workflow Including the QIAseq 1-Step Amplicon Library Kit). It incorporates a one-tube reaction that saves you time, allowing you to focus on sequencing and data analysis. The entire procedure can be performed at room temperature, enabling automation on instruments lacking a thermo-block. The one-tube protocol eliminates the need for transferring reagents, increasing efficiency and more effectively capturing insert amplicons, while also reducing the risk of contamination or sample mix-up, which can occur with manual processing. The procedure is optionally PCR-free to avoid introducing sequence duplicates or PCR-bias, and generates high-quality libraries optimized for sequencing on any Illumina sequencing instrument from a range of input materials.

Excellent sequencing metrics
Typical libraries have excellent sequencing metrics, minimal adapter-adapter ligation product, and specificity and accurately reflect the evenness and sensitivity of the input products. (see figure Typical Results).
Principle
Purified amplicons from gene panels or multiplex PCR are converted to Illumina-compatible NGS libraries using a single, enzymatic library construction step. During this reaction, amplicons are simultaneously prepared for ligation and barcoded adapters are ligated to both ends of the DNA inserts. The adapters contain sequences required for the PCR enrichment of the subsequent library, for flow-cell-binding during bridge amplification and for sequencing primer binding sites for paired-end and multiplexed sequencing.

Following library construction, excess adapters, adapter dimers and other reaction components are removed via precipitation onto Agencourt AMPure XP beads. This procedure is carried out at room temperature, and can be easily automated on various liquid-handling platforms for high-throughput applications. Following library purification, a high-fidelity library enrichment step can be performed to generate sufficient library from low amounts of starting material. This reaction relies on a high-fidelity DNA polymerase and optimized buffer conditions that ensure minimum GC bias and extremely low error rates.

This kit is compatible with:
• PCR products generated with the GeneRead v2 DNAseq Targeted Panels
• PCR products generated with other custom or commercial gene panels
• PCR products generated with the QIAGEN Multiplex PCR Kit or other QIAGEN PCR reagents
• Multiplexed PCR amplicons generated with Taq or Taq derivatives

The novel one-step reaction requires PCR amplicons to contain 3’ A-overhangs for efficient ligation. Taq polymerase, the most commonly used thermostable DNA polymerase, and its derivatives, by default carry out this non-templated A-addition during the PCR reaction. Taq and Taq-derivatives have attributes that make them amenable to multiplex PCR, and many commercial gene panels employ a Taq-based enzyme. In contrast to Taq, other polymerases with strong 3’–5’ exonuclease activities do not carry out this reaction. While these enzymes are not commonly used for multiplexed PCR, amplicons produced with such enzymes are still compatible with the QIAseq 1-Step Amplicon Library Kit, but require A-tailing prior to ligation. Please consult the handbook for details.

Compatible sequencing platforms:
• Illumina HiSeq
• Illumina MiSeq
• Illumina NextSeq
• Illumina MiniSeq
Procedure

The QIAseq 1-Step Amplicon Library Kit offers substantial time savings over standard targeted resequencing library preparations. Utilizing a novel, combined end-repair/ligation reaction, the kit streamlines the entire NGS library preparation process to just 30 minutes. It incorporates a one-tube reaction that saves you time, allowing you to focus on sequencing and data analysis. The entire procedure can be performed at room temperature, enabling automation on instruments lacking a thermo-block. The one-tube protocol eliminates the need for transferring reagents, increasing efficiency and more effectively capturing insert amplicons, while also reducing the risk of contamination or sample mix-up, which can occur with manual processing. The procedure is optionally PCR-free to avoid introducing sequence duplicates or PCR-bias, and generates high-quality libraries optimized for sequencing on any Illumina sequencing instrument from a range of input materials.

Applications

The QIAseq 1-Step Amplicon Library Kit provides an efficient, streamlined, single-reaction solution for amplicon and gene panel sequencing on any Illumina platform.

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