RNA for Illumina®

For use with the Illumina® sequencing platform, mRNA libraries are prepared by construction of a cDNA library, followed by DNA library preparation steps: repair of 3′ and 5′ ends followed by the addition of a non-templated dA-tail before ligation with an adaptor. Libraries are size selected after adaptor ligation, and amplified via polymerase chain reaction (PCR). For more information, choose the mRNA library construction workflow for Illumina tab below.

Small RNA libraries are constructed using a different workflow, in which adaptors are ligated directly to the small RNA molecules, followed by reverse transcription, PCR amplification and size selection. In the novel NEBNext® workflow, the introduction of the RT primer ahead of ligation of the 5’ adaptor prevents formation of contaminating adaptor-dimers. For more information, choose the small RNA library construction workflow for Illumina tab below.

NEBNext reagents are available for each step in mRNA and small RNA library preparation workflows for the Illumina platform. Reagents from NEB are available in a set, master mix or module format. Each of these reagents has been functionally validated by preparation of an RNA library from an accepted RNA reference sample, followed by Illumina sequencing.

Illumina® is a registered trademark of Illumina, Inc.

FAQs for RNA for Illumina®

Protocols for RNA for Illumina®

Ultra II Non-directional RNA Library Preparation Workflow for Illumina

Ultra II Directional RNA Library Preparation Workflow for Illumina

RNA Product Details for Illumina

Small RNA Library Preparation Workflow for Illumina

Small RNA Product Details 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

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.

  1. ABreakthroughMethodOfSmallRNA_720

    A Breakthrough Method of Small 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.