RNA Reagents

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.

  1. 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. The RNA is purified and a fraction of the uncapped RNA is enzymatically capped using the Vaccinia virus capping enzyme. In the final step, the mRNA is transfected into HeLa cells and cell culture supernatants are assayed for luciferase activity.

RNA Reagents Overview

Reagents for RNA Sample Preparation

NEB offers a wide variety of reagents to facilitate front-end processing of RNA for downstream applications including next-generation sequencing, library construction and microarrays.

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.

cDNA Synthesis Selection Chart

NEB offers several reagents for cDNA synthesis for use in applications including qPCR and qRT-PCR. For your convenience, reagents are available as kits or standalone products, depending on your needs.

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.