Next Generation Sequencing

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.
NEBNext® is a registered trademark of New England Biolabs, Inc.

Featured Products

Illumina® Library Preparation includes these areas of focus:

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

FAQs for Illumina® Library Preparation

Protocols for Illumina® Library Preparation

Legal and Disclaimers

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

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.

DNA Library Preparation Workflow for Illumina

RNA Library Preparation Workflow for Illumina

Small RNA Library Preparation Workflow for Illumina