Generate highly complex, Illumina-compatible DNA fragment libraries with ≥50 pg of DNA with this quick and easy kit
- Produces complex libraries with improved sequencing depth uniformity and coverage
- Requires only 50 pg to 75 ng input of sheared/fragmented DNA
- Compatible with any NGS application requiring PCR-amplified libraries
- Minimizes PCR bias to improve coverage uniformity
- Saves time with a fast, <3 hour protocol
The updated NxSeq™ UltraLow DNA library kit v2 combined with a NxSeq™ indexing kit enable the construction of high quality DNA fragment libraries with minimal DNA inputs – as low as 50 pg depending on sample type and illumina sequencer used. If interested in deep sequencing or sequencing more complex genomes/samples, can be used as much as 100 ng of input DNA with this system.
To enable the use of such low input amounts while still generating excellent sequencing results, each step of the protocol is optimized to ensure peak performance on Illumina sequencers. In this updated kit version 2 kit, the original PCR master mix is replaced with a better performing one that is faster with less AT-rich bias. To help with the initial steps of DNA fragmentation, they’ve included 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. Not only high quality libraries will be produced using this kit, but once the fragmented DNA is generated, library prep is quick and easy - a litte under 3 hours.
This system is compatibel with multiple NGS applications including whole genome sequencing (WGS) and resequencing applications (WGR) for de novo sequencing or SNV/mutation or copy number variation (CNV) identification, ChIP-seq, exome-seq and other hybrid capture applications or with challenging sample types such as metagenomic, cell-free and FFPE DNA samples.
To build sequenceable libraries with the NxSeq™ UltraLow DNA library kit, at least one NxSeq™ indexing kit must be purchased. Libraries made with these kits are only compatible with Illumina sequencing instruments (example, NovaSeq, NextSeq, MiSeq and the HiSeq 2500, 3000, 4000, X Ten, and X Five, and iSeq).
Ordering information: The NxSeq™ UltraLow DNA library kit v2, 12 and 96 reactions each contains Enzyme Mix (EM), 2X Buffer (2XB), Ligase (LIG), 2X PCR Master Mix v2 (MM) and Elution Buffer (EB). At least one NxSeq™ indexing kit is required to complete library prep and must be purchased separately.
Each NxSeq™ single indexing kit contains a Universal Adaptor and (12) different primer indexing mixes with enough universal adaptor for 48 libraries and enough of each primer indexing mix for four library amplification reactions (48 total reaction for all primer sets). Set A contains primer indexing mixes 1 to 12 and set B contains primer indexing mixes 13 to 16, 18 to 23, 25, and 27. Each NxSeq™ Single Index equals the TruSeq® LT index with the same number.
Each NxSeq™ HT dual indexing kit contains a universal adaptor, adaptor dilution buffer and indexing primers 701 to 712, and 501 to 508. There is enough of each reagent in this kit to generate 96 different, combinatorial dual indexed libraries. Each NxSeq™ HT Dual Index equals the TruSeq® HT Index with the same number.
Caution: For research use only. Not for human or diagnostic use.
