Product Class: Kit

NEBNext® Ultra™ DNA Library Prep Kit for Illumina®

The small size of this product was discontinued on December 15, 2021. If you would like to purchase a small size, an alternative is NEB #E7645S.

Product Introduction

This original version of the Ultra DNA Library Prep Kit enables library preparation from 5 ng – 1 µg of input DNA in a streamlined workflow. However, we now recommend use of the more recent NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB #E7645), which has been further optimized to provide increased yields of high quality libraries from a broader range of input amounts (500 pg – 1 µg).
Catalog # Size Concentration
E7370L 96.0 reactions

Product Information

Description




The NEBNext® Ultra™ DNA Library Prep Kit for Illumina® contains reagents for preparation of libraries for next-generation sequencing on the Illumina platform from 5 ng – 1 µg input DNA, in a streamlined workflow. Please note that adaptors and primers are not included in the kit and are available separately.

Each kit component must pass rigorous quality control standards, and each set of reagents is functionally validated together by construction and sequencing of a library on the Illumina sequencing platform. 

 

 

Ultra DNA Library Preparation Workflow for IlluminaUltra DNA Library Preparation Workflow for Illumina

NEBNext Ultra™ DNA provides high library yields even with low inputs and difficult samplesNEBNext Ultra™ DNA provides high library yields even with low inputs and difficult samples

All libraries were prepared with 5 ng of input DNA and run on an Agilent Bioanalyzer®.
A. E. coli genomic DNA libraries. 1: Ladder; 2: Library prepared using NEBNext DNA Library Prep Master Mix Set (E6040); 3: Library prepared using NEBNext Ultra DNA Library Prep Kit (E7370). With 5 ng inputs, library yields are significantly higher using the NEBNext Ultra™ DNA Kit.
B. FFPE sample. Human lung tumor genomic DNA from a 66-year old male, extracted from a 10-year old FFPE block (Biochain, Inc.). 1: Library prepared using NEBNext Ultra DNA Library Prep Kit (E7370). 2: Input DNA (highly degraded FFPE genomic DNA).
Read depth correlation shows consistent high coverage for 5 ng - 1 µg input amounts
Read depth correlation shows consistent high coverage for 5 ng - 1 µg input amounts
Libraries were prepared with various amounts of human IMR90 gDNA (5, 50, and 1000 ng) and PCR amplified. A PCR-free library was also prepared with 1000 ng human IMR90 gDNA. Sequence coverage of 10 kb windows of the human genome was analyzed on Galaxy, and Pearson correlation coefficient (R2 values) were calculated using Microsoft® Excel®. All libraries are highly correlated with each other, independent of DNA input as well as PCR amplification. High coverage was achieved for all DNA input amounts.



NEBNext Ultra DNA Kits display strong performance with input amounts as low as 5 ng
NEBNext Ultra DNA Kits display strong performance with input amounts as low as 5 ng

Input DNA: IMR90 genomic DNA (human fibroblast).

PF Mismatch Rate: The rate of bases mismatching the reference for all bases aligned to the reference sequence.

Percent Duplication: The percentage of mapped sequence that is marked as duplicate.

Percent Chimeras: The percentage of reads that map outside of a maximum insert size or that have the two ends mapping to different chromosomes.

This data illustrates the strong performance of the NEBNext Ultra DNA Library Prep Kit for Illumina, even with 5 ng of human genomic DNA input. Mismatch rate, % duplication and % chimeras are all low, and similar, regardless of input amount.

Lot Control

The lots provided are managed separately and qualified by additional functional validation. Individual reagents undergo standard enzyme activity and quality control assays, and also meet stringent criteria in the additional quality controls listed on each individual component page
This product is related to the following categories:

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • E7370L     -20    
      NEBNext End Prep Enzyme Mix E7371AAVIAL -20 1 x 0.288 ml Not Applicable
      NEBNext End Repair Reaction Buffer E7372AAVIAL -20 1 x 0.624 ml 10 X
      Blunt/TA Ligase Master Mix E7373AAVIAL -20 2 x 0.72 ml Not Applicable
      NEBNext® Ligation Enhancer E7374AAVIAL -20 1 x 0.096 ml Not Applicable
      NEBNext Q5 Hot Start HiFi PCR Master Mix E6625AAVIAL -20 2 x 1.2 ml 2 X

Properties & Usage

Materials Required but not Supplied

  • 80% Ethanol (freshly prepared)
  • Nuclease-free Water
  • 0.1X TE, pH 8.0
  • 10 mM Tris-HCl, pH 7.5–8.0 (required for DNA input <100 ng)
  • 5 M NaCl (required for DNA input <100 ng)
  • DNA LoBind Tubes (Eppendorf #022431021)
  • AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
  • NEBNext Singleplex or Multiplex Oligos for Illumina (NEB #E7350, #E7335, #E7500, #E7710, #E7730, #E6609, or #E7600)
  • Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)
  • PCR Machine

Features

  • Input amounts as low as 5ng
  • Fast workflow (<3 hours), with minimal hands-on time
  • Robust, reliable performance
  • Ultra high fidelity amplification with minimized GC bias

Protocols, Manuals & Usage

Protocols

  1. Protocol for use with NEBNext Ultra DNA Library Prep Kit for Illumina (E7370)
  2. Where can I find protocols for using NEBNext® Ultra DNA Library Prep Kit and Modules?
  3. Protocol for use with NEBNext Ultra DNA Library Prep Kit for Illumina (E7370)

Manuals

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

FAQs & Troubleshooting

FAQs

  1. Can AMPure XP beads used for clean-up steps be left in the PCR reactions?
  2. For what type of sample prep/library construction can I use the NEBNext Ultra DNA Library Prep kit for Illumina for?
  3. What type and how much starting material do I need to use when preparing libraries using the NEBNext Ultra DNA Library Prep Kit for Illumina?
  4. Is size selection required and what is the recommended method?
  5. Can I use the NEBNext Ultra DNA Library Prep Kit for Illumina reagents for preparing libraries for sequencing on the Roche 454 GS FLX & SOLiD 5500?
  6. Can I use the NEBNext Ultra DNA Library Prep Kit for Illumina reagents for preparing libraries for sequencing on the ABI SOLiD 4 instrument or on the Ion Torrent PGM?
  7. Can NEB provide any of the reagents for the cluster generation step or the actual sequencing run?
  8. How do I know the concentration of my index primers? 
  9. Which NEBNext Oligos can be used with this library prep kit?
  10. Can I use this NEBNext kit with adaptors and primers from other vendors than NEB?
  11. Are these reagents available in a customized format, larger format or 96 well formats?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

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

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 covered by one or more patents.

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.

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