NxSeq® DNA Sample Prep Kits

Get next-gen library prep in half the time without sacrificing quality.  Next Gen Sequencing (NGS) requires a high performance library preparation kit that drives effective end repair, A-tailing, and adapter ligation. Lucigen's NxSeq® DNA Sample Prep Kits are optimized for highly efficient A-tailing and ligation, ensuring peak performance on your sequencing platform. In addition, the NxSeq kits have one of the most efficient workflows available.  Get sequence ready libraries in less than 2 hours with just a few simple steps.

NxSeq DNA Sample Prep Kits are available for the following platforms:

• Roche 454 (NxSeq DNA Sample Prep Kit 1)
• Illumina (NxSeq DNA Sample Prep Kit 2)
• Ion Torrent – coming soon!

Superior Workflow

Save up to 2 hours off of your current NGS library prep.  NxSeq's optimized master mix format makes the process simpler than ever to create high quality NGS libraries.  Get end-repaired, A-tailed DNA in one simple incubation step, then add ligase and adapters directly to the sample without the need for time consuming and multiple inefficient cleanup steps (see Fig. 1).    Nothing could be easier!

Enhanced Tailing and Ligation

Existing DNA library prep kits do not efficiently A-tail target DNA fragments.  Much of the target DNA is left with blunt ends or even a G-tailed fragment, as Taq and other non-proofreading polymerases add purines at approximately equal rates.  Blunt fragments can ligate with each other, creating chimeras, concatemers, and false junctions.  G-tailed fragments will not ligate with standard T-tailed adapters, resulting in lost data.  NxSeq Technology drives higher efficiency A-tailing and subsequent double-adapter ligation compared to competing kits (see Figure 2). 

Figure 2. Ligation of Texas Red labeled adapter to target DNA.  200 ng of a 90-mer oligo, that cannot be self-ligated without end repair and phosphorylation, was assayed with the NxSeq DNA Sample Prep Kit, the NEBNext™ Quick DNA Sample Prep Master Mix Set 2, and the Roche Library Prep Kit (Rapid Library Rgt/Adaptors).  Tailed reaction product was then ligated to either Texas Red-labeled T-tailed or C-tailed test adaptors to quantify the percentage of target that was either A-tailed or G-tailed.  Product was separated on a 10% acrylamide gel and images were quantitated by gel densitometry of the unstained gel (only Texas Red labeled adaptor signal was quantified).  A)  Ethidium bromide stained gel showing chimera formation. B) Graph of Texas Red signal showing A-tailing and G-tailing efficiencies.

Validated Performance

To validate the NxSeq DNA Sample Prep Kits, Lucigen compared NxSeq vs. Roche, Illumina, and NEB library kits on Roche and Illumina sequencing platforms using E. coli DH10B genomic DNA as a standard.  The result was that Lucigen's NxSeq DNA Sample Prep kits demonstrated equal data quality to that of Roche, NEB or Illumina library prep kits (See Fig. 3 for Illumina comparison). 

Figure 3. Coverage comparison of DH10B Genomic DNA libraries.  Lucigen's NxSeq DNA Sample Prep Kit 2, NEB's NEBNext DNA Sample Prep Master Mix Set 1, and Illumina's TruSeq™ Sample Prep kit-v2 were used to create NGS libraries from DH10B E. coli genomic DNA.  All samples were barcoded and run on an Illumina GAIIx.  Plots indicate depth of coverage achieved in a moving 100 base read window.  Equivalent performance was obtained for all kits.