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

    Description











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

    Figure 1: Workflow
    Figure 1: Workflow.
    Functional Validation
    Each set of reagents is functionally validated together through construction and sequencing of a DNA library on the Illumina sequencing platform.

    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 Universal PCR Primer for Illumina-20
    NEBNext Index 1 Primer for Illumina-20
    NEBNext Index 2 Primer for Illumina-20
    NEBNext Index 3 Primer for Illumina-20
    NEBNext Index 4 Primer for Illumina-20
    NEBNext Index 5 Primer for Illumina-20
    NEBNext Index 6 Primer for Illumina-20
    NEBNext Index 7 Primer for Illumina-20
    NEBNext Index 8 Primer for Illumina-20
    NEBNext Index 9 Primer for Illumina-20
    NEBNext Index 10 Primer for Illumina-20
    NEBNext Index 11 Primer for Illumina-20
    NEBNext Index 12 Primer for Illumina-20

    Properties and Usage

    Materials Required but not Supplied

    Enzymes and buffers appropriate for DNA or ChIP Illumina library preparation. (DNA: #E7645 , #E7370 , #E6040 , #E6000     ChIP: #E7645 , #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 Tris-HCl, pH 8.0
    • 10 mM NaCl
    • Bioanalyzer® (Agilent Technologies, Inc.)
    • Bio-Rad® Microseal® (Bio-Rad Laboratories, Inc.) "A" Film or equivalent
    Materials Required but not supplied 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 ), or NEBNext rRNA Depletion Kit (Human, Mouse, Rat) (NEB #E6310 ) or similar
    • 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
    • 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 (Agilent Technologies, Inc.)
    • Bio-Rad Microseal (Bio-Rad Laboratories, Inc.) "A" Film or equivalent
    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)
    Please Refer to the Kit Specific Protocol for using the NEBNext Multiplex Oligos for Illumina:
    The following kits are designed for use with the NEBNext Multiplex Oligos for Illumina:
    • #E7645 , NEBNext Ultra II DNA Library Prep Kit for Illumina
    • #E7595 , NEBNext Ultra II Ligation Module
    • #E7420 , NEBNext Ultra Directional RNA Library Prep Kit for Illumina
    • #E7530 , NEBNext Ultra RNA Library Prep Kit for Illumina
    • #E7370 , NEBNext Ultra DNA Library Prep Kit for Illumina
    • #E7445 , NEBNext Ultra Ligation Module
    • #E6040 , NEBNext DNA Library Prep Master Mix Set for Illumina
    • #E6110 , NEBNext mRNA Library Prep Master Mix Set for Illumina
    • #E6240 , NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina
    • #E6056 , NEBNext Quick Ligation Module
    • #E6000 , NEBNext DNA Library Prep Reagent Set for Illumina
    • #E6200 , NEBNext ChIP-Seq Library Prep Reagent Set for Illumina
    • #E6100 , NEBNext mRNA Library Prep Reagent Set for Illumin

    Storage Temperature

    -20°C

    FAQs

    1. Are NEBNext adaptor and primers for Illumina validated in Next Generation Sequencing Workflows?
    2. Are NEBNext adaptor and primers for Illumina compatible with NEBNext reagents for Illumina library preparation?
    3. Is USER enzyme required for library preparation using NEBNext adaptor and primers for Illumina?
    4. How much DNA can I ligate to the NEBNext adaptor for Illumina in one ligation reaction?
    5. Are these reagents available in a customized format? Larger format or 96 well format?
    6. Can USER Enzyme be purchased separately?
    7. Are the NEBNext Oligos for Illumina compatible with single end and paired end sequencing?
    8. Is the USER enzyme in our NEBNext kits identical in concentration to the standalone product?
    9. How do I know the concentration of my index primers? 

    Protocols

    1. Please see manual (NEB #E7335) for instructions.

    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].

    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.