QIAseq and GeneRead Library Prep Kits

For fast and efficient preparation of DNA libraries for use in NGS applications

Features

  • Fast, one-tube procedure that allows up to 50% time savings
  • High yields from minimal amounts of starting material
  • Unbiased amplification with an optional, high-fidelity amplification step
  • Multiplexing, high-throughput capability
GeneRead DNA Library I Core Kit (48)

Cat. No. / ID: 180434

For 48 reactions: Buffers and reagents for end-repair, A-Addition, and ligation, for use with Illumina instruments
KitAccessories
DNA Library Core Kit
Adapter Set
QIAseq 1-Step Amplicon Library Kit
FX DNA Library Kit
Ultralow Input Library Kit
DNA Library Kit
DNA Amp Kit
For instrument
Illumina
Ion Torrent
Reactions
48
12
QIAseq and GeneRead Library Prep Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

GeneRead Library Prep Kits provide an optimized and efficient workflow for library preparation in next-generation sequencing (NGS) applications. The kits allow fast and efficient preparation of DNA libraries that are for use on NGS platforms from Illumina and the Ion Torrent instrument from Life Technologies. Procedures can be automated on the QIAcube.


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Performance

High yields of library DNA with uniform coverage distribution
GeneRead Library Prep Kits provide an efficient and optimized workflow to reproducibly generate high yields of DNA library, with minimal sequence bias and low error rates, for use on NGS platforms from Illumina or the Ion Torrent instrument from Life Technologies. Highly efficient ligation reactions and an optional, high-fidelity amplification step ensure superior library yields with uniform coverage distribution, even from as low as 50 ng starting material (see figures  High yields of library DNA with uniform coverage distribution and  Superior yields of library DNA).

Low error rates with minimal sequence bias
To ensure maximum yields from minimum amounts of starting material, GeneRead Library Prep Kits include an innovative high-fidelity master mix that includes GeneRead HiFi Polymerase for an optional amplification step. GeneRead HiFi Polymerase is a unique, highly specific and processive enzyme that delivers accurate amplification of library DNA with low error rates and minimum bias (see figure  Low error rates with minimal sequence bias due to high-fidelity amplification).

Lower sequence bias when amplifying GC- and AT-rich regions
In standard PCR amplification procedures, regions of DNA with high AT or GC content can result in little or no amplification, leading to misleading sequence data and NGS results. The GeneRead HiFi Polymerase, together with its unique buffer formulation, ensures uniform amplification of genomic regions that contain highly variable GC content, thereby ensuring even coverage in subsequent sequencing reactions (see figure  Better sequence coverage).
See figures

Principle

GeneRead Library Prep Kits use an optimized, one-tube workflow to reproducibly generate high yields of DNA library for use on NGS platforms from Illumina or the Ion Torrent instrument from Life Technologies. The fast, one-tube procedure uses fewer cleanup steps than library preparation workflows from other suppliers and includes an optional, high-fidelity library amplification step. GeneRead Library Prep Kits save time and effort while minimizing the variability caused by handing, as well as the risk of contamination (see figures  Optimized workflow for Illumina-compatible DNA libraries and  Optimized workflow for Ion Torrent-compatible DNA libraries).
See figures

Procedure

Procedure for generating Illumina-compatible DNA libraries
Samples consisting of longer DNA fragments are first sheared into a random library of fragments that are a median fragment size of 300 bp (when using the Illumina MiSeq instrument, version 1), or 500 bp (when using the Illumina MiSeq instrument, version 2), in length. Following fragmentation, the ends of the DNA fragments are repaired and an A-overhang is added at the 3'-end of each strand. Afterwards, adaptors, which are necessary for amplification and sequencing, are ligated to both ends of the DNA fragments. Barcode adapters, which contain a unique identifying sequence, are also available with the GeneRead Library Prep (I) Kit and enable multiplex sequencing reactions to be performed. Fragments are then size selected and purified. Cleanup and size selection can be done using the GeneRead Size Selection Kit (cat. no. 180514), which allows precise, column-based size selection of DNA (see figure  Precise size selection). To ensure maximum yields from minimum amounts of starting material, an optional high-fidelity amplification step can also be performed.

Procedure for generating Ion Torrent-compatible DNA libraries
Samples consisting of longer DNA fragments are first sheared into a random library of fragments that are a median fragment size of 400 bp (when using the Ion Torrent PGM instrument), or 200 bp (when using the Ion Proton instrument), in length. Following fragmentation, the ends of the DNA fragments are repaired. Afterwards, adaptors, which are necessary for amplification and sequencing, are ligated to both ends of the DNA fragments. Barcode adapters, which contain a unique identifying sequence, are also available with the GeneRead Library Prep (L) Kit and enable multiplex sequencing reactions to be performed. Fragments are then size selected and purified. To ensure maximum yields from minimum amounts of starting material, an optional high-fidelity amplification step can be performed.

See figures

Applications

GeneRead Library Prep Kits allow fast and efficient preparation of DNA libraries that are optimized for use on NGS platforms from Illumina and the Ion Torrent instrument from Life Technologies.

Supporting data and figures

Resources

Kit Handbooks (5)
For preparation of DNA libraries for next-generation sequencing (NGS) applications that use Illumina instruments
For preparation of DNA libraries for next-generation sequencing (NGS) applications that use Ion Torrent instruments from Life Technologies
Additional Resources (1)
For the preparation, size selection, and purification of DNA libraries for NGS applications