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RNA-seq

Next generation sequencing (NGS) can be used to determine the presence and quantity of RNA species in a sample enabling sensitive and accurate gene expression analysis. For the commonly used 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).

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.

Libraries are then clonally amplified and sequenced using sequencing by synthesis (sbs) methods, such as the Illumina sequencing platform.

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.


FAQs for RNA-seq

Protocols for RNA-seq

    Other Tools & Resources

    Brochures

  • Latest NEB Expressions

    This edition of NEB expressions focuses on tools that can streamline your RNA-related workflows. Featured products include NEBNext Ultra II kits for RNA, the Monarch Total RNA Miniprep Kit, Luna reagents for RT-qPCR, the new LunaScript RT SuperMix Kit and HiScribe kits for high yield, high quality RNA synthesis. Learn more about functional considerations for RNA synthesis in our feature, and learn about a new protocol for Cappable Seq. Lastly, read about ways you can promote sustainable science in the lab.

  • RNA Synthesis Brochure

    Learn more about NEB's products for RNA synthesis, which range from template generation to poly(A) tailing.

  • RNA Technical Guide

    Find in depth information on tools designed to streamline your RNA workflows, including RNA extraction & purification, qPCR and RT-qPCR, RNA-seq, RNA synthesis and gRNA synthesis.

  • Usage Guidelines

  • Getting Started with RNA-Sequencing (RNA-Seq)

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 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.

  1. NEBNextUltra2DirectionalRNA_720

    NEBNext Ultra II Directional RNA Workflow

    Learn more about the streamlined workflow for the NEBNext Ultra II Directional RNA Library Prep Kit.

  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.