NEBNext^® Ultra^™ II DNA Library Prep with Sample Purification Beads

Product Information

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.

Advantages

• 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

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

For use with NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors DNA Set 1) (NEB https://www.neb.com/en/#E7395), refer to the Protocols tab for UMI Adaptors-specific guidance. 

This product is related to the following categories:

DNA Library Prep for Illumina,

Analysis of DNA Methylation by Bisulfite Conversion,

Automation for NEBNext® NGS Library Prep Products,

DNA Methylation Analysis

• Kit Components

  The following reagents are supplied with this product:

  NEB https://www.neb.com/en/#	Component Name	Component https://www.neb.com/en/#	Stored at (°C)	Amount	Concentration
  E7103S     Multi-temperature       NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® E7645S -20       NEBNext® Ultra II End Prep Enzyme Mix E7646AVIAL -20 1 x 0.072 ml Not Applicable   NEBNext® Ultra II End Prep Reaction Buffer E7647AVIAL -20 1 x 0.168 ml Not Applicable   NEBNext® Ultra™ II Ligation Master Mix E7648AVIAL -20 1 x 0.72 ml Not Applicable   NEBNext® Ligation Enhancer E7374AVIAL -20 1 x 0.024 ml Not Applicable   NEBNext® Ultra II Q5® Master Mix E7649AVIAL -20 1 x 0.6 ml Not Applicable   NEBNext® Sample Purification Beads E7104S 25       NEBNext® Sample Purification Beads E7104SVIAL 25 1 x 4 ml Not Applicable
  E7103L     Multi-temperature       NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® E7645L -20       NEBNext® Ultra II End Prep Enzyme Mix E7646AAVIAL -20 1 x 0.288 ml Not Applicable   NEBNext® Ultra II End Prep Reaction Buffer E7647AAVIAL -20 1 x 0.672 ml Not Applicable   NEBNext® Ultra™ II Ligation Master Mix E7648AAVIAL -20 3 x 0.96 ml Not Applicable   NEBNext® Ligation Enhancer E7374AAVIAL -20 1 x 0.096 ml Not Applicable   NEBNext® Ultra II Q5® Master Mix E7649AAVIAL -20 2 x 1.2 ml Not Applicable   NEBNext® Sample Purification Beads E7104L 25       NEBNext® Sample Purification Beads E7104SVIAL 25 4 x 4 ml Not Applicable

• Properties & 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 https://www.neb.com/en/#022431021) 

  • NEBNext Multiplex Oligos for Illumina (for a full list, visit www.neb.com/oligos)

  • Magnetic rack or plate (e.g., NEBNext^® Magnetic Separation Rack (NEB https://www.neb.com/en/#S1515S), Alpaqua ^® 96S Super Magnet Plate (https://www.neb.com/en/#A001322), or equivalent)
  • PCR Machine

  Optional:

  • 10 mM Tris-HCl, pH 8.0 or 
  • Tris/NaCl, pH 8.0 (10 mM Tris, 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.

Protocols, Manuals & Usage

• Protocols

  1. Where can I find protocols for using NEBNext^® DNA Library Prep reagents?

• Manuals

  The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

  • manualE7103-E7645
  • manualE7103_E7645 w UMI Adaptors DNA

• Application Notes

  • Improved Library Preparation with NEBNext^® Ultra™ II DNA Library Prep Kit for Illumina^®

FAQs & Troubleshooting

• FAQs

  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, larger format or 96 well formats?
  7. I am currently using Agencourt Ampure XP beads. Will the NEBNext Sample Purification Beads be a suitable replacement?
  8. Which NEBNext Oligos can be used with this library prep kit?
  9. Can I use this NEBNext kit with adaptors and primers from other vendors than NEB?
  10. What do I do if I see a precipitate in the Ultra II End Prep Reaction Buffer?

• Tech Tips

  When removing the supernatant from the magnetic beads during cleanup, itis important not to disturb the pellet. Find these and other helpfultips for optimizing the size selection and clean up steps in thisprotocol video.
   

  

Citations & Technical Literature

• Citations

  Product Citation Tool

Quality, Safety & Legal

• Quality Control Assays

  Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

• Safety Data Sheets

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

  • NEBNext^® Ultra^™ II DNA Library Prep with Sample Purification Beads
  • NEBNext^® Ultra II End Prep Enzyme Mix
  • NEBNext^® Ultra II End Prep Reaction Buffer
  • NEBNext^® Ultra^™ II Ligation Master Mix
  • NEBNext^® Ligation Enhancer
  • NEBNext^® Ultra II Q5^® Master Mix

• Legal and Disclaimers

  Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
  
  This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
  
  New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

  Licenses

  This product is licensed for research and commercial use from Bio-Rad Laboratories, Inc., under U.S. Pat. Nos. 6,627,424, 7,541,170, 7,670,808, 7,666,645, and corresponding patents in other countries. No rights are granted for use of the product for Digital PCR or real-time PCR applications, with the exception of quantification in Next Generation Sequencing workflows.

  For additional information or to inquire about commercial use, please contact busdev@neb.com.