GeneRead DNA I Amp Kit 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

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.

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.

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 .

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 .

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 .

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. 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.

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.