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  • NEBNext® Ultra™ Directional RNA Second Strand Synthesis Module


    The NEBNext® Ultra™ Directional RNA Second Strand Synthesis Module has been optimized to generate double-stranded cDNA from first-strand cDNA. The dsDNA generated by the NEBNext Ultra Directional RNA Second Strand Module can be subsequently converted to blunt-end DNA fragments using the NEBNext Ultra End Prep Module.

    The NEBNext Ultra Directional RNA Second Strand Synthesis Module is Designed for Use with the Following:
    NEBNext RNA First Strand Synthesis Module (NEB #E7525)
    NEBNext Ultra™ End Repair/dA-Tailing Module (NEB #E7442)
    NEBNext Ultra™ Ligation Module (NEB #E7445)
    NEBNext Singleplex or Multiplex Oligos for Illumina® (NEB #E7350, #E7335 or #E7500)
    USER™ Enzyme (NEB #M5505) Note: This is included in the NEBNext Oligo sets; however, if you are using alternative oligos, it is necessary to purchase separately for the directional method.
    NEBNext High-Fidelity 2X PCR Master Mix (NEB #M0541)

    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 Second Strand Synthesis Enzyme Mix-20
    NEBNext Second Strand Synthesis Reaction Buffer with dUTP Mix-2010X

    Properties and Usage

    Storage Temperature



    1. What is the difference between the NEBNext Ultra Directional RNA Second Strand Module and the NEBNext mRNA Second Strand Synthesis Module?
    2. For the downstream workflow, which Modules for End Repair, dA-Tailing and Ligation should be used with the NEBNext Ultra Directional Second Strand Synthesis Module?


    1. Protocol (E7550)


    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.