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In vitro Synthesis (IVT)

In vitro synthesis of single-stranded RNA molecules is a widely used laboratory procedure that is critical to RNA research, as well as to RNA biopharmaceuticals. This technique is versatile in that it allows the researcher to tailor synthesis and introduce modifications to produce a transcript. Downstream applications include biochemical and molecular characterization of RNA for RNA-protein interactions and structural analyses, generation of RNA aptamers, synthesis of functional mRNAs for expression, and generation of small RNAs for alteration of gene expression (e.g., guide RNAs, RNAi). Furthermore, the use of in vitro synthesized RNA has been instrumental in the development of RNA vaccines and CRISPR/Cas9 genome editing tools, generation of pluripotent stem cells, as well as development of RNA amplification-based diagnostics.

High-yield robust IVT reactions require optimization of each reaction component. NEB offers five in vitro RNA synthesis kits, all of which have been optimized to generate reproducible yields of quality RNA. In addition, individual components can also be purchased for IVT and mRNA capping.

  1. Rio, D. C., et al. RNA: A Laboratory Manual. Cold Spring Harbor: Cold Spring Harbor Laboratory Press, 2011, 205-220.
  2. Cooper, Geoffery M. The Cell: A Molecular Approach. 4th ed. Washington D.C.: ASM Press, 2007. 262-299.
  3. Jani B, Fuchs R. J Vis Exp. 2012 Mar 26 (61). PMID: 22473375

FAQs for In vitro Synthesis (IVT)

Protocols for In vitro Synthesis (IVT)

    Other Tools & Resources

    Brochures

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  • RNA Synthesis Brochure

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    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.

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  • Usage Guidelines

  • Avoiding Ribonuclease Contamination

Recommended HiScribe RNA Synthesis Kits by Application

RNA Polymerase Selection Chart

NEB offers RNA polymerases that can be used for in vitro synthesis of RNA for a wide variety of downstream applications. Additional polymerases are offered for the generation of untemplated homoribopolymeric tails for reverse transcription or labeling.

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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    In Vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection

    This method describes high yield in vitro synthesis of both capped and uncapped mRNA from a linearized plasmid containing the Gaussia luciferase (GLuc) gene.