Product Class: Other

NEBNext® RNA First Strand Synthesis Module

This product was discontinued on December 15, 2022. An alternative product is the NEBNext® Ultra™ II RNA First Strand Synthesis Module (NEB #E7771S).

  • Catalog # E7525 was discontinued on December 15, 2022

Product Introduction

This module is optimized for use in the original Ultra™ workflow, for Illumina®-compatible RNA library construction. The original Ultra workflow enables library preparation from 5 ng – 1 µg of input DNA in a streamlined protocol.

However, we now recommend use of the more recent NEBNext Ultra II Directional RNA and RNA (non-directional) workflows, which have been further optimized to provide increased yields of high quality libraries from a broader range of input amounts, and a NEBNext Ultra II RNA First Strand Synthesis Module (NEB #E7771) is available.


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Catalog # Size Concentration

Product Information

Description



The NEBNext RNA First Strand Synthesis Module is designed for synthesis of cDNA from RNA fragments using ProtoScript® II Reverse Transcriptase and random priming. Single strand cDNA can be used directly for second strand cDNA synthesis. For use in directional (strand-specific) protocols, Actinomycin D is required and must be purchased separately (see below).
This product is related to the following categories:

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • E7525S     -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
      ProtoScript® II Reverse Transcriptase E7423AVIAL -20 1 x 0.024 ml Not Applicable
  • E7525L     -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
      ProtoScript® II Reverse Transcriptase E7423AAVIAL -20 1 x 0.096 ml Not Applicable

Properties & Usage

Materials Required but not Supplied

If you are using the module for Directional RNA Seq, Actinomycin D (Sigma #A1410 dissolved in dimethylsulfoxide [DMSO] to 5μg/μl) is required. See protocol for details.

Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)

Protocols, Manuals & Usage

Protocols

  1. Protocol for Use with Total RNA - NEBNext mRNA First Strand Synthesis Module (E7525)
  2. Protocol for Use with Previously Fragmented RNA - NEBNext mRNA First Strand Synthesis Module (E7525)

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 other materials do I need in addition to the NEBNext First Strand Synthesis Module?
  2. What if I have already fragmented my mRNA and just need to perform the First Strand Synthesis Reaction?

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.

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