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.
|= 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.
- A Typical DNase I Reaction Protocol (M0303)
- Capped RNA Synthesis (E2040)
- Capping Protocol (M2080)
- DNA Template Preparation (E2040)
- Evaluation of Reaction Products (E2040)
- High Specific Activity Radiolabeled RNA Probe Synthesis (E2040)
- A Typical Tailing Reaction (M0337)
- Purification of Synthesized RNA (E2040)
- RNA Synthesis with Modified Nucleotides (E2040)
- SP6 In Vitro Transcription Reaction Protocol (M0207)
- Standard RNA Synthesis (E2040)
- 2´-O-Methylation of Capped RNA (M0366)
- One-Step Capping and 2´-O-Methylation (M0366)
- Labeling Protocol (M2080)
- Capped RNA Synthesis (E2050)
- Evaluation of Reaction Products (E2050)
- Purification of Synthesized RNA (E2050)
- RNA Synthesis with Modified Nucleotides (E2050)
- Standard RNA Synthesis (E2050)
- In vitro digestion of DNA with Cas9 Nuclease, S. pyogenes (M0386)
- Poly(A) Tailing of RNA using E. coli Poly(A) Polymerase (NEB# M0276)
- Protocol for Standard RNA Synthesis
- Evaluation of Reaction Products (E2060)
- mRNA Purification (E2060)
- mRNA Purification (E2065)
- mRNA Synthesis with Modified Nucleotides (E2060)
- mRNA Synthesis with Modified Nucleotides (E2065)
- Standard mRNA Synthesis (E2060)
- Standard mRNA Synthesis (E2065)
- Evaluation of Reaction Products (E2065)
- Using recombinant Cas9 nuclease to assess locus modification in genome editing experiments (#M0386)
- sgRNA Synthesis Using the HiScribe™ Quick T7 High Yield RNA Synthesis Kit (NEB #E2050)
- EnGen® sgRNA Synthesis Kit, S. pyogenes Protocol (E3322)
- Transfection of Cas9 RNP (ribonucleoprotein) into adherent cells using the Lipofectamine® RNAiMAX
- Purification of Synthesized RNA (E2070)
- RNA Synthesis Protocols (E2070)
- Transfection of EnGen® Spy Cas9 HF1 (NEB #M0667) into adherent cells using the Lipofectamine® RNAiMAX System
- Avoiding Ribonuclease Contamination
- 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
- Wu, M.Z., Asahara, H., Tzertzinis, G., Roy, B. (2020) Synthesis of low immunogenicity RNA with high-temperature in vitro transcription RNA; PubMedID: 31900329
- 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
- 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
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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.
Interested in GMP-grade IVT reagents?
This method describes high yield in vitro synthesis of both capped and uncapped mRNA from a linearized plasmid containing the Gaussia luciferase (GLuc) gene.