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  • NEBNext® DNA Library Prep Reagent Set for Illumina®


    The DNA Library Prep Reagent Set for Illumina 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.

    DNA Library Preparation Workflow for Illumina

    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
    Phosphorylation Reaction Buffer-2010X
    Deoxynucleotide Solution Mix-2010 mM
    T4 DNA Polymerase-20
    DNA Polymerase I, Large (Klenow) Fragment-20
    T4 Polynucleotide Kinase-20
    Deoxyadenosine 5´- Triphosphate (dATP)-201 mM
    Klenow Fragment (3´→ 5´ exo)
    NEBuffer 2 for Klenow Fragment (3´→ 5´ exo)10X
    Quick T4 DNA Ligase-20
    NEBNext® Quick Ligation Reaction Buffer-202X
    NEBNext® Q5® Hot Start HiFi PCR Master Mix-20
    NEBNext High-Fidelity 2X PCR Master Mix (SAMPLE)2X

    Properties and Usage

    Materials Required but not Supplied

    • 80% Ethanol (freshly prepared)
    • Nuclease-free Water
    • 0.1X TE, pH 8.0
    • 10 mM Tris-HCl, pH 7.5–8.0 (optional)
    • DNA LoBind Tubes (Eppendorf #022431021)
    • AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
    • NEBNext Singleplex or Multiplex Oligos for Illumina (NEB #E7350, #E7335, #E7500 or #E7600)
    • Magnetic rack/stand
    • PCR Machine

    Storage Temperature



    1. For larger volume requirements, customized and bulk packaging is available by purchasing through the OEM/Bulks department at NEB. Please contact OEM@neb.com for further information.


    1. What type of sample prep/library construction can I use the NEBNext DNA Library Prep Reagent Set for Illumina reagents for?
    2. What type and how much starting material do I need to use when preparing libraries using the NEBNext DNA Library Prep Reagent Set for Illumina?
    3. Can I use the NEBNext DNA Library Prep Reagent Set for Illumina reagents for preparing RNA? For miRNA work?
    4. Can I use these reagents for preparing libraries for sequencing on the ABI SOLiD instrument?
    5. Can NEB provide any of the reagents for the cluster generation step or the actual sequencing run?
    6. Can I use the NEBNext DNA Library Prep Reagent Set for Illumina reagents for preparing libraries for sequencing on the Roche 454 GSFLX?
    7. Are these reagents available in a customized format? Larger format or 96 well format?
    8. Do I need to dA-tail my insert for cloning an expression library?
    9. What are the differences between expression library preparation vs. sample preparation suitable for the Illumina platform?


    1. Please see manual (NEB #E6000) for protocols.


    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].

    Safety Data Sheet

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.