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RNA Library Prep

NEBNext® Ultra™ II kits are available for directional (strand-specific) or non-directional RNA library preparation. These kits utilize streamlined workflows and novel master mixes, and have been designed for performance with input amounts as low as 5 ng Total RNA.

Modules offer the user ability to customize sample preparation, and adaptors and primers (NEBNext Oligos) are available separately, for maximum flexibility.

  • The NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (NEB #E7760) uses the dUTP method and a streamlined workflow to generate high yield, high quality strand-specific libraries.
  • The NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (NEB #E7770) enables production of non-directional libraries from as little as 10 ng of Total RNA, using a streamlined workflow.

For microRNAs and other small RNAs, our novel Small RNA workflow has been optimized to minimize adaptor-dimers, while producing high-yield, high-density libraries.

For single cell and ultra-low input RNA, our simple and robust workflow generates full length cDNAs, followed by conversion to sequence-ready libraries using the Ultra II FS workflow, that incorporates enzymatic DNA fragmentation.

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FAQs for RNA Library Prep

Protocols for RNA Library Prep

Other Tools and Resources

Single Cell/Low Input RNA Library Preparation Workflow for Illumina


Ultra II Directional RNA Library Preparation Workflow for Illumina


Ultra II  Non-directional 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

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. NEBNextVideo_OptimizingRNAInput_thumb

    Tips for Optimizing RNA Inputs in NGS Library Construction

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

  2. 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.

  3. NEBNextVideo_BeadBasedCleanup_thumb

    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.

  4. NEBNextVideo_12Tips_thumb

    12 Quick Tips for NGS Library Preparation

    These 12 quick tips for NGS library preparation are a great refresher if you're a seasoned pro, or a great introduction if you're a beginner.