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  • RNA Modules

    NEB offers a variety of modules for use in the preparation of RNA for the leading sequencing platforms:

    NEBNext® Poly(A) mRNA Magnetic Isolation Module

    NEBNext® Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) is optimized to isolate intact poly(A)+ RNA from previously isolated total RNA. The module couples Oligo d(T)25 and 1 μm paramagnetic beads for the direct binding of poly(A)+ RNA. The procedure permits the manual processing of multiple samples and can be adapted for automated high-throughput applications. Additionally, magnetic separation technology permits elution of intact mRNA in small volumes eliminating the need for precipitating the poly(A)+ transcripts in the eluent.

    NEBNext® Magnesium RNA Fragmentation Module

    NEBNext® Magnesium RNA Fragmentation Module (NEB #E6150) is optimized to fragment RNA into small pieces using divalent cations under elevated temperature. The module contains buffers that are suited for RNA fragmentation.

    NEBNext® RNase III RNA Fragmentation Module

    NEBNext® RNase III RNA Fragmentation Module (NEB #E6146) is 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 module contains enzyme and buffer that are suited for RNA fragmentation.

    NEBNext® Second Strand Synthesis Module

    NEBNext® Second Strand Synthesis Module (NEB #E6111) is optimized to generate double stranded cDNA from first strand cDNA. The module is provided as a master mix to maximize efficiency and convenience in RNA sample preparation workflows. The dsDNA generated by the NEBNext Second Strand Synthesis Module (NEB #E6111) can be subsequently converted to blunt-ended DNA fragments using the NEBNext End Repair Module (NEB #E6050).

    Advantages of NEBNext Reagents for RNA Library Preparation

    • Convenient Formats – Enzymes, buffers and nucleotides are included, at appropriate concentrations and in appropriate volumes, and are available in set or master mix format. Large and small sizes are available. 
    • Functional Validation – Each set is validated together by preparation of a library from an accepted RNA reference sample, followed by sequencing on the appropriate sequencing platform (Illumina, 454 or SOLiD). 
    • Stringent Quality Controls
    • Value Pricing
    • Novel Protocol for small RNA – Higher yield and substantially reduced adaptor dimer formation suitable for methylated and unmethylated small RNAs

    Applications of NEBNext Reagents for RNA Library Preparation

    • mRNA library preparation for next generation sequencing
    • Small RNA library preparation for sequencing with Illumina and SOLiD
    • Expression library construction

    mRNA Library Construction Workflow

    mRNA Library Construction Workflow for Illumina

    mRNA Library Construction Workflow for 454

    mRNA Library Construction Workflow for SOLiD