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  • NEBNext® mRNA Library Prep Set for SOLiD™

    • This product was discontinued on 01/01/2013
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    Discontinued Products

    Description

    The NEBNext® mRNA Library Prep Set for SOLiD™ contains enzymes and buffers that are ideally suited for sample preparation for next-generation sequencing, and for preparation of expression libraries. Each of these components must pass rigorous quality control standards and are Lot Controlled, both individually and as a set of reagents.

    Figure 1: Relative size distribution of eukaryotic mRNA fragments as seen using the Bioanalyzer 2100.
    Figure 2: Bioanalyzer traces of mRNA Fragments with insufficient washing show a noisy baseline.
    Figure 3: Final mRNA-Seq Library Bioanalyzer Trace
    Functional Validation
    Each set of reagents is functionally validated together through construction and sequencing of an RNA library on the SOLiD™ 4.

    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 RNA Fragmentation Buffer10X
    NEBNext RNA Fragmentation Stop Solution10X
    Linear Acrylamide-2010 mg/ml
    Random Primers
    Murine RNase Inhibitor
    NEBNext First Strand Synthesis Reaction Buffer5X
    NEBNext Second Strand Synthesis Enzyme Mix
    NEBNext Second Strand Synthesis Reaction Buffer10X
    Nuclease-free Water-20
    NEBNext End Repair Reaction Buffer10X
    T4 DNA Ligase
    NEBNext Quick Ligation Reaction Buffer-205X
    LongAmp Taq 2X Master Mix2X
    NEBNext End Repair Enzyme Mix-20

    Properties and Usage

    Materials Required but not Supplied

    • 3 M Sodium Acetate, pH 5.5 
    • 100% Ethanol 
    • 70% Ethanol 
    • SuperScript® II Reverse Transcriptase (Life Technologies, Inc.)
    • PCR Column Purification Kit (Qiagen or other) 
    • DNA Gel Extraction Column Purification Kit 
    • DNA Adaptors and Primers or Vector
    • Size Selection Materials [E-Gel® (Life Technologies, Inc.), Agarose Gel or AMPure XP Beads]

    Storage Temperature

    -20°C

    FAQs

    1. Do you provide a protocol for using these reagents?
    2. What type of sample prep/library construction can I use these Sets for?
    3. How much starting material do I need to use when preparing libraries using the NEBNext® RNA sets?
    4. Do I need any reagents that are not supplied in the kit?

    Protocols

    1. mRNA Fragmentation Protocol (E6114)
    2. First Strand cDNA Synthesis (E6114)
    3. Second Strand cDNA Synthesis (E6114)
    4. NEBNext End Repair cDNA Library (E6114)
    5. Adaptor or Vector Ligation of cDNA Library (E6114)
    6. PCR Amplification of Adaptor Ligated cDNA Library (E6114)
    7. First Strand cDNA Synthesis

    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.