GeneRead DNA Library I Kit
- 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 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 canbe automated on theQIAcube.
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Cat No./ID:180435 GeneRead DNA Library I Kit (96) $2,908.00 Buffers and reagents for end-repair, A-addition, ligation and library amplification; for use with Illumina instruments; includes a plate containing 96 adapters with different barcodes (pierceable foil seal allowing usage of defined parts of plate) |
Product Details
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 Polymerasefor an optional amplification step. GeneRead HiFi Polymeraseisa unique, highly specific and processive enzyme thatdelivers 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).
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 fromother suppliers and includesan optional, high-fidelity library amplificationstep. GeneRead Library Prep Kitssave time and effort while minimizingthe variability caused by handing, as well asthe risk of contamination (see figuresOptimized workflow for Illumina-compatible DNA libraries and Optimized workflow for Ion Torrent-compatible DNA libraries).
Procedure for generating Illumina-compatible DNA libraries
Samples consisting of longer DNA fragments are firstsheared into a random library of fragments that are a median fragment size of 300 bp (when using theIllumina 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 firstsheared 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.
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.
