Many next generation DNA sequencing applications or sample types require the construction of PCR-amplified DNA fragment libraries. (E.g. whole genome sequencing or resequencing from limiting genomic DNA amounts or FFPE and cell-free DNA samples, exome sequencing, ChIP-seq, etc.). To get the highest quality data for these applications and sample types, you need a DNA library preparation kit with the highest efficiency adaptor ligation followed by unbiased PCR amplification to produce the most complex, unbiased libraries possible. These high quality libraries produce the high quality sequencing data with uniform depth and minimal zero coverage needed to drive your NGS experiments forward.
The updated NxSeq™ UltraLow DNA Library Kits v2 and NxSeq Indexing Kits allow you to build high quality DNA fragment libraries from extremely low DNA input amounts – as low as 50 pg depending on sample type and Illumina sequencer used. If you have more DNA, no problem; you can use as much as 100 ng of input DNA with this system.
To generate these high quality libraries, each step of this protocol has been optimized to ensure peak performance on Illumina sequencers. To help with the initial steps of DNA fragmentation, included is guidance for mechanical shearing on an instrument like a Covaris LE220 and an optimized protocol for enzymatic fragmentation using dsDNA Shearase Plus from Zymo Research. In this updated kit, they replaced the PCR master mix with an improved master mix that performs better by decreasing bias across high AT-rich genomic regions. Not only will you produce high quality libraries, but also once you have your fragmented DNA, library prep is quick and easy - about three total hours.
These library and indexing kits are compatible with multiple applications and sample types including whole genome sequencing (WGS) and resequencing (WGR) applications for genomic assembly, SNV/mutation or copy number variation (CNV) identification. The system is also compatible with other NGS applications such as ChIP-seq and exome-seq. It also works well with both high quality input DNA samples as well as challenging sample types such as metagenomic, FFPE and cell-free DNA samples.
