Product Class: Kit

NEBNext® Ultra™ RNA Library Prep Kit for Illumina®

This product will be discontinued on December 15, 2022. An alternative product is the NEBNext® Ultra™ II RNA Library Prep Kit for Illumina®  (NEB #E7770).

  • Catalog # E7530 was discontinued on December 15, 2022

Product Introduction

This original version of the Ultra™ RNA Library Prep Kit enables non-directional RNA library preparation in a streamlined workflow. However, we now recommend use of the more recent NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB #E7770), which has been further optimized to provide increased yields of high quality libraries from a broader range of input amounts. 
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Catalog # Size Concentration

Product Information

Description

 

The NEBNext® Ultra™ RNA Library Prep Kit for Illumina® contains reagents for preparation of non-directional RNA libraries for Illumina sequencing. Please note that adaptors, primers, rRNA depletion reagents and poly(A) mRNA isolation reagents 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. 

NEBNext Ultra™ RNA provides even sequence coverage across transcripts for a wide range of inputs
NEBNext Ultra™ RNA provides even sequence coverage across transcripts for a wide range of inputs

10 ng, 100ng or 1 μg of Total RNA (Universal Human Reference RNA, Agilent #740000) was used as input for libraries, following the NEBNext Ultra™ RNA protocol and using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490). Libraries were sequenced (single end reads) on the Illumina platform (GAIIx). Good coverage across expressed transcript was obtained for each input amount.

Ultra Non-directional RNA Library Preparation for IlluminaNon-Directional

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
  • E7530S     -20    
      NEBNext First Strand Synthesis Reaction Buffer E7421AVIAL -20 1 x 0.192 ml Not Applicable
      Random Primers E7422AVIAL -20 1 x 0.048 ml Not Applicable
      Murine RNase Inhibitor E7424AVIAL -20 1 x 0.015 ml Not Applicable
      NEBNext Second Strand Synthesis Enzyme Mix E7425AVIAL -20 1 x 0.096 ml Not Applicable
      NEBNext End Prep Enzyme Mix E7371AVIAL -20 1 x 0.072 ml Not Applicable
      NEBNext End Repair Reaction Buffer E7372AVIAL -20 1 x 0.156 ml 10 X
      Blunt/TA Ligase Master Mix E7373AVIAL -20 1 x 0.36 ml Not Applicable
      Nuclease-free Water E7431AVIAL -20 1 x 8 ml Not Applicable
      NEBNext Second Strand Synthesis Reaction Buffer E7427AVIAL -20 1 x 0.192 ml Not Applicable
      ProtoScript® II Reverse Transcriptase E7423AVIAL -20 1 x 0.024 ml Not Applicable
      NEBNext Q5 Hot Start HiFi PCR Master Mix E6625AVIAL -20 1 x 0.6 ml 2 X
  • E7530L     -20    
      NEBNext First Strand Synthesis Reaction Buffer E7421AAVIAL -20 1 x 0.768 ml Not Applicable
      Random Primers E7422AAVIAL -20 1 x 0.192 ml Not Applicable
      Murine RNase Inhibitor E7424AAVIAL -20 1 x 0.048 ml Not Applicable
      NEBNext Second Strand Synthesis Enzyme Mix E7425AAVIAL -20 1 x 0.384 ml Not Applicable
      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
      Nuclease-free Water E7431AAVIAL -20 1 x 30 ml Not Applicable
      NEBNext Second Strand Synthesis Reaction Buffer E7427AAVIAL -20 1 x 0.768 ml Not Applicable
      ProtoScript® II Reverse Transcriptase E7423AAVIAL -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

  • NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) or NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310)
  • NEBNext Singleplex (NEB #E7350) or NEBNext Multiplex (NEB #E7335, #E7500 or #7600) Oligos for Illumina
  • Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)
  • 80% Ethanol (freshly prepared)
  • Agencourt® AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
  • Agencourt RNAClean® XP (Beckman Coulter, Inc. #A63987) are required when using NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310)
  • 0.1X TE, pH 8.0
  • 10mM Tris-HCl, pH 7.5–8.0
  • 10 mM NaCl (optional)

Features

  • Low input amounts (as low as 10ng total RNA, purified mRNA or ribosomal-depleted RNA)
  • Fast workflow (5 – 5.5 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® Poly(A) mRNA Magnetic Isolation Module (E7490) and NEBNext® Ultra RNA Library Prep Kit for Illumina® (E7530)
  2. Protocol for use with NEBNext rRNA Depletion Kit (Human/Mouse/Rat), NEB #E6310 and NEBNext Ultra RNA Library Prep Kit for Illumina (NEB #E7530)
  3. Protocol for use with Purified mRNA or Ribosome Depleted RNA - NEBNext Ultra RNA Library Prep Kit  for Illumina (E7530)

Manuals

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

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. What is the difference between the NEBNext Ultra Directional RNA library prep kit for Illumina (E7420) and the NEBNext Ultra RNA library prep kit for Illumina (E7530)?
  2. What is the starting material I need to use when preparing libraries using the NEBNext Ultra Directional RNA kit?
  3. Where do I have to start the protocol if I have purified mRNA or Ribosomal-depleted RNA?
  4. Which kit can I use to isolate Poly (A) mRNA from Total RNA?
  5. Does the kit provide adaptor and primers?
  6. How do I know the concentration of my index primers? 
  7. Which NEBNext Oligos can be used with this library prep kit?
  8. Can I use this NEBNext kit with adaptors and primers from other vendors than NEB?

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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