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.

GenomeWeb article reviews NEBNext Small RNA Kits.


Illumina® is a registered trademark of Illumina, Inc.

Illumina® Library Preparation includes these areas of focus:

RNA for Illumina®
DNA for Illumina®
NEBNext® Ultra™ II DNA Library Prep
NEBNext® Ultra™ II RNA Library Prep

FAQs for Illumina® Library Preparation

Protocols for Illumina® Library Preparation

    Other Tools & Resources


  • 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.

  • The NEBNext Sample Preparation Workflow Poster

    The NEBNext Sample Preparation Workflow Overviews & Products poster gives you at-a-glance workflow details for our DNA & RNA library prep products for Illumina & Ion Torrent, as well as our latest featured products.

DNA Library Preparation Workflow for Illumina

RNA Library Preparation Workflow for Illumina

Small RNA Library Preparation Workflow for Illumina

Legal Information

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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