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  • NEBNext® Multiplex Oligos for Illumina® (Dual Index Primers Set 1)

    Description






    The NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1) contains the adaptor and index primers that are ideally suited for multiplex sample preparation for next-generation sequencing on the Illumina platform. Each of these components must pass rigorous quality control standards and are lot controlled, both individually and as a set of reagents.

    Dual Index Workflow

    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 Adaptor for Illumina-20
    USER Enzyme-20
    NEBNext i501 Primer-20
    NEBNext i502 Primer-20
    NEBNext i503 Primer-20
    NEBNext i504 Primer-20
    NEBNext i505 Primer-20
    NEBNext i506 Primer-20
    NEBNext i507 Primer-20
    NEBNext i508 Primer-20
    NEBNext i701 Primer-20
    NEBNext i702 Primer-20
    NEBNext i703 Primer-20
    NEBNext i704 Primer-20
    NEBNext i705 Primer-20
    NEBNext i706 Primer-20
    NEBNext i707 Primer-20
    NEBNext i708 Primer-20
    NEBNext i709 Primer-20
    NEBNext i710 Primer-20
    NEBNext i711 Primer-20
    NEBNext i712 Primer-20

    Properties and Usage

    Materials Required but not Supplied

    Required Materials Not Included for DNA or ChIP Libraries:
    • Enzymes and buffers appropriate for DNA or ChIP Illumina library preparation. (DNA: #E7370, #E6040, #E6000 ChIP: #E7370, #E6240, #E6200)
    • Agencourt® AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
    • Magnetic Stand
    • DNA LoBind® Tubes (Eppendorf®) / PCR Tubes/96-well PCR Plates
    • Freshly prepared 80% Ethanol
    • Nuclease-free Water
    • 0.1X TE (or 10 mM Tris-HCl, pH 7.5–8.0)
    • 10 mM NaCl (optional) Bioanalyzer® (Agilent Technologies, Inc.)
    • Bio-Rad® Microseal® (Bio-Rad Laboratories, Inc.) "A" Film or equivalent
    • Recommended products not supplied:
      TruSeq® Index Plate Fixture (Illumina #FC-130-1005) or similar setup
    Required Materials Not Included for RNA Libraries:
    • Enzymes and buffers appropriate for RNA Illumina library preparation. (#E7420, #E7530, #E6110, #E6100)
    • NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490)
    • Magnetic Rack (Alpaqua, cat #A001322 or equivalent)
    • 80% Ethanol (freshly prepared)
    • 0.1X TE (or 10 mM Tris-HCl, pH 7.5–8.0)
    • 10 mM NaCl (optional)
    • Agencourt AMPure XP Beads (Beckman Coulter, Inc. #A63881)
    • Actinomycin D (Sigma# A1410, dissolved in dimethylsulfoxide [DMSO] to 5 μg/µl).
    • DNA LoBind Tubes/PCR Tubes/96-well PCR Plates
    • Bioanalyzer
    • Bio-Rad Microseal "A" Film or equivalent
    • Recommended products not supplied:
      TruSeq Index Plate Fixture (Illumina #FC-130-1005) or similar setup.
    • Additional materials required for use with #E6110 and #E6100:
      3 M Sodium Acetate, pH 5.5 100% Ethanol PCR Column Purification Kit (Qiagen or other) DNA Gel Extraction Column Purification Kit RNeasy® MinElute® Clean Up Kit (Qiagen® #74204)

    Storage Temperature

    -20°C

    FAQs

    1. Do NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1)come with library prep reagents?
    2. Can I use NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1) for library prep from both DNA and RNA samples?
    3. How many libraries can I prepare with the NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1)?
    4. For pooling <96 samples, what combination of indices should I use?
    5. Do I need to spike in custom sequencing primers when sequencing libraries made with NEBNext Multiplex Oligos for Illumina (Dual Index Primers Set 1) on the Illumina platforms?
    6. Can I perform single read runs and still get both index sequences? *
    7. Can I run both single read and paired-end recipes with dual-indexed libraries? *
    8. How does dual indexing work? *
    9. Is the USER enzyme in our NEBNext kits identical in concentration to the standalone product?

    Protocols

    1. Setting up the PCR Reaction (E7600)

    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.