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    NEBNext® Small RNA Library Prep Set for SOLiD™

    Description

    The NEBNext Small RNA Library Prep Set for SOLiD™ contains adaptors, primers, enzymes and buffers that are ideal to convert small RNA into cDNA libraries for next-generation sequencing on the SOLiD platform. NEBNext Small RNA Library Prep Set for Solid is multiplex compatible. Each of these components must pass rigorous quality control standards and are lot controlled, both individually and as a set of reagents.

    Functional Validation
    Each set of reagents is functionally validated together through construction and sequencing of an RNA library on the SOLiD™ 4.

    Recent article in GenomeWeb reviews NEBNext Small RNA Kits.

    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 3´ Ligation Reaction Buffer-202X
    NEBNext 3´ SR Adaptor 3-20
    NEBNext 5´ SR Adaptor 3-20
    NEBNext 5´ Ligation Reaction Buffer-20
    NEBNext SR RT Primer 3-20
    Murine RNase Inhibitor
    NEBNext SR Primer R3-20
    Quick-Load pBR322 DNA-MspI Digest-2050 μg/ml
    DNA Gel Elution Buffer-201X
    Linear Acrylamide10 mg/ml
    TE Buffer
    Nuclease-free Water-20
    OneTaq Hot Start 2X Master Mix with Standard Buffer-202X
    NEBNext First Strand Synthesis Reaction Buffer-205X
    NEBNext Index 1 Primer for SOLiD
    ProtoScript® II Reverse Transcriptase-20200,000 units/ml
    NEBNext 3´ Ligation Enzyme Mix-20
    NEBNext 5´ Ligation Enzyme Mix-20
    Gel Loading Dye, Blue (6X)25

    Properties and Usage

    Materials Required but not Supplied


    3 M Sodium Acetate, pH 5.2
    100% Ethanol
    80% Ethanol
    SuperScript® II Reverse Transcriptase (Invitrogen #18064-014)
    5X First Strand Buffer (Supplied with SuperScript® II Reverse Transcriptase)
    0.1 M DTT (Supplied with SuperScript® II Reverse Transcriptase)
    Costar Spin-X centrifuge tube Filters (Cellulose Acetate Filters) (Sigma #CLS8162)
    RNase-free Disposable Pellet Pestles (Kontes #74951-1590)
    6% Novex TBE PAGE gel, 1.0 mm, 10 well (Invitrogen #EC6265BOX)
    SYBR® Gold Nucleic Acid Gel Stain (Invitrogen #S-11494)

    Storage Temperature

    -20°C

    FAQs

    1. Is Index 1 in the NEBNext Small RNA Library Prep Set for SOLiD kit (E6160) compatible with the NEBNext Small RNA Multiplex (1-16) Prep Set for SOLiD kit (E7450)?
    2. If my input is <5ug and therefore use 0.5 ul of  3' SR Adapter 3, SR RT Primer 3 and 5' SR Adapter 3, instead of 1.0 ul, my total volumes for each reaction will be short by 0.5 ul. Should I replace that lost volume with water?
    3. Why do I add the SR RT Primer 3 before the 5’ ligation instead of right before the reverse transcription step?
    4. If I forget to add the 3’ SR RT Primer 3 before the 5’ ligation can I add it before the reverse transcription step and move forward?
    5. Can samples be stored longer than 72 hours at -20°C?
    6. How many cycles of PCR should I do?
    7. If I accidentally move all 30 ul of the 5’ ligation product forward to the reverse transcription mixture will the reaction still work?

    Protocols

    1. Library Preparation Using NEBNext® Small RNA Library Prep Set for SOLiD™ (E6160)

    Manuals

    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].

    Material Safety Data Sheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.