NEBNext® RNA First Strand Synthesis Module


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.

Functional Validation

Each set of reagents is functionally validated together through construction and sequencing of a transcriptome library and sequenced on a .

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

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
NEBNext First Strand Synthesis Reaction Buffer-20
Random Primers-20
ProtoScript® II Reverse Transcriptase-20200,000 units/ml
Murine RNase Inhibitor-20

Properties and Usage

Materials Required but not Supplied

NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490)

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.

Storage Temperature



  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?


  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)


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].

Usage Guidelines & Tips

Interactive Tools

Safety Data Sheet

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