Next Generation Sequencing
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  • Illumina® Library Preparation

    Library preparation of fragmented DNA, ChIP-DNA, and cDNA prepared from mRNA, for use with the Illumina® sequencing platform, involves repair of 3’ and 5’ ends followed by the addition of a non-templated dA-tail before ligation to an adaptor, and PCR amplification. Small RNA libraries are constructed using a different workflow, in which adaptors are ligated directly to the small RNA molecules, followed by reverse transcription and PCR amplification. For more information, choose the workflow tabs below.

    Libraries prepared in this manner are suitable for cluster generation, the next step in the Illumina sequencing workflow.

    NEBNext® reagents are available for each step in the library preparation workflow for the Illumina platform. For DNA, ChIP, mRNA and small RNA library preparation for sequencing with the Illumina series of instruments, reagents from NEB are available in a set, master mix set or module format. Each of these reagents has been functionally validated by preparation of a genomic DNA, ChIP-Seq DNA, mRNA or small RNA library, followed by Illumina sequencing.

    Recent article in GenomeWeb reviews NEBNext Small RNA Kits.

    Illumina® is a registered trademark of Illumina, Inc.
    NEBNext® is a registered trademark of New England Biolabs, Inc.
    • Why Choose NEBNext Reagents for NGS Library Prep?

      NEBNext products are a great choice for your NGS library preparation needs. In this video, Fiona shares a few reasons why.

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    • 12 Quick Tips for NGS Library Preparation

      Using beads to clean up your DNA prior to NGS library prep can be quick and easy, if you follow these few, simple tips.

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    • Tips for Optimizing RNA Inputs in NGS Library Construction

      Watch as Deyra explains how to optimize your RNA inputs for successful NGS library preparation.

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    • NEBNext for Core Lab Facilities

      Are you running a core sequencing facility? Watch as Eileen shares a few reasons that NEBNext products are particularly well suited to sequencing core labs.

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    • Which NEBNext Product Format Should I Choose?

      Not sure how sets and kits differ from modules? Ping explains the difference, and helps you to decide which one is right for you.

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    • Tips for Bead-based Clean ups and Size Selection

      Using beads to clean up your DNA prior to NGS library prep can be quick and easy, if you follow these few, simple tips.

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    • A Breakthrough Method of RNA Sample Preparation

      The common problem of adaptor dimer formation during small RNA library construction can be avoided by using NEBNext® protocols. Learn about this technique, and how it improves both performance and sensitivity in library construction.

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    • Tips for Optimizing DNA Inputs in NGS Library Construction

      Optimizing DNA inputs for NGS library prep is an important step, if you want to ensure a high-quality library. Start by following these tips.

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    Featured Products

    Illumina® Library Preparation includes these areas of focus:

    DNA for Illumina®
    RNA for Illumina®

    FAQs for Illumina® Library Preparation

      Other Tools & Resources

      Brochures

    • NEBNext® for Illumina® Brochure

      The NEBNext Brochure for Illumina provides an overview of library preparation for DNA, ChIP-Seq, RNA and Small RNA for the Illumina platform, as well as tech tips and select peer-reviewed citations.

    DNA Library Construction Workflow for Illumina

    mRNA Library Construction Workflow for Illumina

    Small RNA Library Construction Workflow for Illumina