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  • NEBNext® Multiplex Small RNA Library Prep Set for SOLiD™ (Set 1)

    Description

    The NEBNext Multiplex Small RNA Library Prep Set for SOLiD (Set 1) contains adaptors, primers, enzymes and buffers that are ideal to convert small RNA into cDNA libraries for next-generation sequencing on the SOLiD platform (Life Technologies, Inc.). Each of these components must pass rigorous quality control standards and are lot controlled, both individually and as a set of reagents.

    Small RNA Library Prep Workflow
    Novel Protocol that Results in Higher Yields and Lower Adaptor-dimer Contamination (Patent Pending):
    E7450
    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-2010X
    NEBNext SR RT Primer 3-20
    NEBNext First Strand Synthesis Reaction Buffer-205X
    ProtoScript® II Reverse Transcriptase-20200,000 units/ml
    Murine RNase Inhibitor-20
    OneTaq Hot Start 2X Master Mix with Standard Buffer-202X
    NEBNext Index 1 Primer for SOLiD-20
    NEBNext Index 2 Primer for SOLiD-20
    NEBNext Index 3 Primer for SOLiD-20
    NEBNext Index 4 Primer for SOLiD-20
    NEBNext Index 5 Primer for SOLiD-20
    NEBNext Index 6 Primer for SOLiD-20
    NEBNext Index 7 Primer for SOLiD-20
    NEBNext Index 8 Primer for SOLiD-20
    NEBNext Index 9 Primer for SOLiD-20
    NEBNext Index 10 Primer for SOLiD-20
    NEBNext Index 11 Primer for SOLiD-20
    NEBNext Index 12 Primer for SOLiD-20
    NEBNext Index 13 Primer for SOLiD-20
    NEBNext Index 14 Primer for SOLiD-20
    NEBNext Index 15 Primer for SOLiD-20
    NEBNext Index 16 Primer for SOLiD-20
    NEBNext SR Primer R3-20
    Gel Loading Dye, Blue256X
    Quick-Load pBR322 DNA-MspI Digest-20100 μg/ml
    DNA Gel Elution Buffer41X
    Linear Acrylamide-2010 mg/ml
    TE Buffer-20
    Nuclease-free Water-20
    NEBNext 3´ Ligation Enzyme Mix-20
    NEBNext 5´ Ligation Enzyme Mix-20

    Properties and Usage

    Materials Required but not Supplied

    • 3M Sodium Acetate. pH 5.2
    • 100% Ethanol
    • 80% Ethanol 
    • Corning®, Costar®, Spin-X® Centrifuge Tube Filters (Cellulose Acetate Filters)(Sigma Aldrich, #CLS8162)
    • RNase-free Disposable Pellet Pestles® (Kimble Kontes Asset Management, Inc., #749521-1590) 
    • 6% Novex® TBE PAGE gel, 1.0 mm, 10 well (Life Technologies, Inc., E-C6265BOX) 
    • SYBR® Gold Nucleic Acid Gel Stain (Life Technologies, Inc., S-11494)

    Storage Temperature

    -20°C

    Notes

    1. Kit Optimization:
      The components of this kit were collectively optimized with FirstChoice® Human Brain Reference RNA (Life Technologies, Inc. #AM6050).

    Supporting Documents

    Material Safety Datasheets

    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.

    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].
    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?
    1. Library Preparation Using NEBNext® Multiplex Small RNA Library Prep Set for SOLiD™ (Set 1) (E7450)