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.
- Evaluation of Reaction Products (E2065)
- mRNA Synthesis with Modified Nucleotides (E2065)
- Standard mRNA Synthesis (E2060)
- Standard mRNA Synthesis (E2065)
- mRNA Synthesis with Modified Nucleotides (E2060)
- Evaluation of Reaction Products (E2060)
- mRNA Purification (E2060)
- mRNA Purification (E2065)
- Protocol for Standard RNA Synthesis
- RNA Synthesis Protocols (E2070)
- Purification of Synthesized RNA (E2070)
- Capped RNA Synthesis (E2040)
- High Specific Activity Radiolabeled RNA Probe Synthesis (E2040)
- DNA Template Preparation (E2040)
- Evaluation of Reaction Products (E2040)
- RNA Synthesis with Modified Nucleotides (E2040)
- Purification of Synthesized RNA (E2040)
- Standard RNA Synthesis (E2040)
- Capped RNA Synthesis (E2050)
- Evaluation of Reaction Products (E2050)
- Purification of Synthesized RNA (E2050)
- RNA Synthesis with Modified Nucleotides (E2050)
- Standard RNA Synthesis (E2050)
- sgRNA Synthesis Using the HiScribe™ Quick T7 High Yield RNA Synthesis Kit (NEB #E2050)
- EnGen® sgRNA Synthesis Kit, S. pyogenes Protocol (E3322)
- SP6 In Vitro Transcription Reaction Protocol (M0207)
- 2´-O-Methylation of Capped RNA (M0366)
- Capping Protocol (M2080)
- Labeling Protocol (M2080)
- One-Step Capping and 2´-O-Methylation (M0366)
- A Typical DNase I Reaction Protocol (M0303)
- Transfection of Cas9 RNP (ribonucleoprotein) into adherent cells using the Lipofectamine® RNAiMAX
- A Typical Tailing Reaction (M0337)
- In vitro digestion of DNA with Cas9 Nuclease, S. pyogenes (M0386)
- Using recombinant Cas9 nuclease to assess locus modification in genome editing experiments (#M0386)
- Poly(A) Tailing of RNA using E. coli Poly(A) Polymerase (NEB# M0276)
- Avoiding Ribonuclease Contamination
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 [email protected].
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.
This method describes high yield in vitro synthesis of both capped and uncapped mRNA from a linearized plasmid containing the Gaussia luciferase (GLuc) gene.