NEBNext® Ultra II DNA Library Prep with Sample Purification Beads


The NEBNext Ultra II DNA Library Prep with Sample Purification Beads contains the enzymes, buffers and purification beads required to convert a broad range of DNA input amounts into indexed libraries for next-generation sequencing on the Illumina platform. The fast, user-friendly workflow has minimal hands-on time and, for added convenience, this kit contains NEBNext Sample Purification Beads (SPRIselect® beads from Beckman Coulter) for size selection and enzyme reaction cleanup.

  • Get more of what you need, with the highest library yields
  • Use to generate high quality libraries even when you have only limited amounts of DNA, with inputs as low as 500 pg
  • Prepare libraries from ALL of your samples, including GC-rich targets and FFPE DNA samples
  • Improve yields and quality for target enrichment applications
  • Save time with streamlined workflows, reduced hands-on time and automation compatibility, and enjoy the flexibility of kit or module formats
  • Enjoy the flexibility and reliability of the gold standard SPRIselect size selection and clean-up beads, supplied in just the amounts you need
For extensive NEBNext Ultra II performance data, click the links in the Advantages above or visit the NEBNext Ultra II DNA Library Prep Kit for Illumina page.

To learn more about how NEBNext Ultra II addresses lower input amounts and challenging sample types, download our technical note.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
NEBNext Ultra II End Prep Enzyme Mix-20
NEBNext Ultra II End Prep Reaction Buffer-20
NEBNext® Ultra II Ligation Master Mix-20
NEBNext® Ligation Enhancer-20
NEBNext® Ultra II Q5® Master Mix-202X
NEBNext Sample Purification Beads25

Properties and Usage

Materials Required but not Supplied

  • 80% Ethanol (freshly prepared) 
  • Nuclease-free Water 
  • 0.1X TE (1 mM Tris-HCl, pH 8.0, 0.1 mM EDTA) 
  • DNA LoBind Tubes (Eppendorf #022431021) 
  • NEBNext Singleplex or Multiplex Oligos for Illumina (E7350, E7335, E7500, E6609, E7600, E7100, E7730
  • Magnetic rack/stand 
  • PCR Machine
  • 10 mM Tris-HCl, pH 8.0 or 
  • 10 mM Tris-HCl, pH 8.0 with 10 mM NaCl (for adaptor dilution of DNA input ≤ 100 ng)

Storage Temperature:
Package 1: NEBNext Ultra II DNA Library Prep Kit for Illumina - Store at -20°C
Package 2: NEBNext Sample Purification Beads - Store at Room Temperature. Do not freeze.


  1. For what type of sample prep/library construction can I use the NEBNext Ultra II DNA Library Prep kit for Illumina?
  2. What type and how much starting material do I need to use when preparing libraries using the NEBNext Ultra II DNA Library Prep Kit for Illumina?
  3. Is size selection required and what is the recommended method?
  4. Can I use the NEBNext Ultra II DNA Library Prep Kit for Illumina reagents for preparing libraries for sequencing on the Ion Torrent PGM or Proton?
  5. Can NEB provide any of the reagents for the cluster generation step or the actual sequencing run?
  6. Are these reagents available in a customized format? Bulk or 96 well formats?
  7. I am currently using Agencourt Ampure XP beads. Will the NEBNext Sample Purification Beads be a suitable replacement?

Tech Tips

When removing the supernatant from the magnetic beads during cleanup, it is important not to disturb the pellet. Find these and other helpful tips for optimizing the size selection and clean up steps in this protocol video.


The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name these document files: manual[Catalog Number].

Application Notes

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.