RNA Synthesis

Tools to Take You from Template to Transcript

For almost 50 years, NEB® has been a world leader in the discovery and production of reagents for the life science industry. New England Biolabs’ enzymology expertise allows us to supply reagents for the synthesis of high-quality RNA – from template generation and transcription, to capping, tailing, and cleanup after synthesis. These products are designed and manufactured by building upon decades of molecular biology experience, so that you can be confident they will work for your application.

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, screening of RNA inhibitors, as well as development of RNA amplification-based diagnostics.

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

NEB’s portfolio of research-grade and GMP-grade* reagents enables bench-scale to commercial-scale mRNA manufacturing. Our optimized HiScribe kits enable convenient in vitro transcription workflows. When it is time to scale up and optimize reaction components, our standalone reagents are readily available in formats matching our GMP-grade offering, enabling a seamless transition to large-scale therapeutic mRNA manufacturing.


Template Generation TableCaret_GMPgrade In vitro Transcription TableCaret_GMPgrade RNA Capping TableCaret_GMPgrade Poly(A) Tailing TableCaret_GMPgrade RNAPurification
Q5® High-Fidelity DNA Polymerase HiScribe T7 mRNA Kit with CleanCap® Reagent AG E. coli Poly(A) Polymerase Monarch® RNA Cleanup Kit (10 μg)
HiScribe®; T7 ARCA mRNA Synthesis Kit (with tailing)
dNTP solution mixes HiScribe T7 ARCA mRNA Synthesis Kit Monarch RNA Cleanup Kit (50 μg)
BspQI*   GMPgrade_icon HiScribe T7 High Yield RNA Synthesis Components   Faustovirus Capping Enzyme GMP_ComingSoon_Icon_42w   Monarch RNA Cleanup Kit (500 μg)
Vaccinia Capping System GMPgrade_icon

DNA Assembly:

HiScribe T7 Quick High Yield RNA Synthesis Kit   mRNA Cap 2′-O-Methyltranferase GMPgrade_icon   Lithium Chloride
  HiScribe SP6 High Yield RNA Synthesis Kit   ARCA and other mRNA cap analogs      
  T3 & SP6 RNA Polymerases          
  T7 RNA Polymerase  GMPgrade_icon        

Hi-T7 RNA Polymerase

  Companion Products         Companion Products
  RNase inhibitor (Murine)  GMPgrade_icon       Monarch RNA Cleanup Binding Buffer
  RNase Inhibitor (Human Placental)         Monarch RNA Cleanup Wash Buffer
  Pyrophosphatase, Inorganic (E. coli)  GMPgrade_icon       Nuclease-free Water
  Pyrophosphatase, Inorganic (Yeast)          
  DNase I (RNase-free)  GMPgrade_icon        
  DNase I-XT

GMPgrade_icon = available in GMP-grade

*NEB can offer large-scale preparations of restriction enzymes using Recombinant Albumin (BSA-free)

"GMP-grade" is a branding term NEB uses to describe reagents manufactured at NEB’s Rowley facility. The Rowley facility was designed to manufacture reagents under more rigorous infrastructure and process controls to achieve more stringent product specifications and customer requirements. Reagents manufactured at NEB’s Rowley facility are manufactured in compliance with ISO 9001 and ISO 13485 quality management system standards. However, at this time, NEB does not manufacture or sell products known as Active Pharmaceutical Ingredients (APIs), nor does NEB manufacture its products in compliance with all of the Current Good Manufacturing Practice regulations. 

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Protocols for RNA Synthesis
    Publications related to RNA Synthesis
    • Wu, M.Z., Asahara, H., Tzertzinis, G., Roy, B. (2020) Synthesis of low immunogenicity RNA with high-temperature in vitro transcription RNA; PubMedID: 31900329
    • Wulf, Madalee; Buswell, John; Chan, Siuhong; Dai, Nan; Marks, Katherine; Tzertzinis, George; Whipple, Joe; Correa, Ivan; Schildkraut, Ira; (2019) The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping Sci Rep; 9 (1), 8594. PubMedID: 31197197, DOI: 10.1038/s41598-019-45083-5
    • Eyler, D.E., Franco, M.K., Batool, Z., Wu, M.Z., Dubuke, M.L., Dobosz-Bartoszek, M., Jones, J.D., Polikanov, Y.S., Roy, B., Koutmou, K.S (2019) Pseudouridinylatio of mRNA coding sequences alters translation Proc Natl Acad Sci U S A; 116(46), 23068-23074.. PubMedID: 31672910, DOI: 10.1073/pnas.1821754116
    • Potapov, V., Fu, X., Dai, N., Correa, I.R., Jr., Tanner, N.A., Ong, J.L (2018) Base modificatons affecting RNA polymerase and reverse transcriptase fidelity Nucleic Acids Res; 46(11): 5753-5763, PubMedID: 29750267
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.
Cap Analog Selection Chart
NEB offers a number of cap analogs that increase stability, decrease susceptibility to exonuclease degradation, and promote the formation of mRNA initiation complexes.
Legal Information

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@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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