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  • NEBNext® RNase III RNA Fragmentation Module


    The NEBNext RNase III RNA Fragmentation Module has been optimized to fragment RNA into small pieces using RNase III enzyme. RNase III is an endonuclease enzyme that cleaves long double-stranded RNA into short fragments containing 5´-PO4 and 3´-OH ends.

    The NEBNext RNase III RNA Fragmentation Module contains enzyme and buffer that are suited for RNA fragmentation. Each of these components must pass rigorous quality control standards listed on each individual component page.

    Figure 1: Relative size distribution of eukaryotic mRNA fragments as seen using the Bioanalyzer 2100. Poly (A)+ mRNA (100 nanograms) purified from the Universal Human Reference Total RNA (Stratagene) was fragmented in 1X NEBNext RNase III RNA Reaction Buffer for 5 minutes at 37°C using different amounts of the NEBNext RNase III enzyme (1; 0.5; 0.25; 0.125; 0.062; 0.031 units). RNA fragments were column purified and analyzed on the Bioanalyzer 2100.

    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 RNase III-201 units/μl
    NEBNext RNase III Reaction Buffer-2010X

    Advantages and Features


    RNA Fragmentation

    Properties and Usage

    Materials Required but not Supplied

    RNeasy MinElute Cleanup Kit (Qiagen # 74204)
    3M Sodium Acetate, pH 5.2
    100% Ethanol
    70% Ethanol
    Linear Acrylamide 10 mg/ml

    Storage Temperature



    1. What are the differences between the reaction products of fragmentation with the NEBNext® Magnesium RNA Fragmentation Module and the NEBNext® RNase III RNA Fragmentation Module?
    2. Which kit should I use for fragmentation of my RNA?
    3. How long should I incubate the fragmentation reaction?
    4. How much enzyme should I use in the fragmentation reaction?
    5. How can I purify the RNA fragments?
    6. Do I need any reagents that are not supplied in the kit?
    7. How can I assess the yield and the size distribution of the fragmented RNA?
    8. Which sizes of RNA fragments are recommended for Next Generation Sequencing?


    1. NEBNext® RNase III RNA Fragmentation Module (E6146)


    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

    Safety Data Sheet

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