RNA Synthesis

RNA Polymerases

T7 (NEB # M0251) and SP6 (NEB # M0207) RNA polymerases are DNA dependent RNA polymerases that produce DNA templated RNA transcripts. T7 and SP6 exhibit high specificity for their respective promoters. Both T7 and SP6 can be used for the in vitro synthesis of RNA for a wide variety of applications, including transfection, translation, structural studies and radioactive and non-isotopic probe generation. E. coli Poly(A) Polymerase (NEB # M0276) and Poly(U) Polymerase (NEB # M0337) generate untemplated homoribopolymeric tails on the 3´-ends of RNA. Both E. coli Poly(A) Polymerase and Poly(U) Polymerase can be used for RNA tailing for reverse transcription or labeling.

In vitro RNA synthesis requires a DNA template, RNA polymerase, NTPs and other factors. High-yield robust reactions require optimization of each reaction component. NEB offers five in vitro RNA synthesis kits, all of which have been rigorously formulated to provide reproducible high yields of quality RNA. Additionally, NEB offers a Vaccinia capping system (NEB # M2080) and a number of RNA cap analogs.

Learn more about the HiScribe in vitro transcription kits.

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Protocols for RNA Synthesis
    Publications related to RNA Synthesis
  1. 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 Scientific Reports. 9 (1), PubMedID: , DOI: 10.1038/s41598-019-45083-5
  2. 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 USA. 116(46), PubMedID: 31672910, DOI: 10.1073/pnas.1821754116
  3. Wu, M.Z., Asahara, H., Tzertzinis, G., Roy, B. 2020. Synthesis of low immunogenicity RNA with high-temperature in vitro transcription . , PubMedID: 31900329, DOI:
  4. 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 Research. 46(11): 5753-5763, PubMedID: 29750267, DOI:
RNA Reagents Overview
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.
RNA Ligase Selection Chart
NEB offers a variety of ligases for RNA research with unique specificities.
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.
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