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RNA Synthesis Kits

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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 optimized to generate reproducible yields of quality RNA.

For efficient translation to occur, most eukaryotic mRNAs require a 7-methyl guanosine (m7G) cap structure at the 5′ end and a poly(A) tail at the 3′ end. The HiScribe™ T7 ARCA mRNA Synthesis Kit (NEB #E2060S) is designed to synthesize capped and tailed mRNAs for variety of applications. Capped mRNAs are synthesized by co-transcriptional incorporation of Anti-Reverse Cap Analog, ARCA, using T7 RNA Polymerase. A poly(A) tail is then added by E. coli Poly(A) Polymerase. This kit is also available without E. coli Poly(A) Polymerase (NEB #E2065S) for use with DNA templates encoding a poly(A) stretch or not requiring a poly(A) tail. Both kits include DNase I and LiCl for DNA template removal and quick mRNA purification.

The HiScribe T7 High Yield RNA Synthesis Kit (NEB #E2040) delivers robust RNA synthesis for a wide range of template sizes. Flexible protocols ensure that performance is maintained, even under demanding conditions, such as extended reaction time using very low amounts of template. Protocols are included for partial or complete incorporation of modified or labeled nucleotides in the transcript body, and cap analogs at the RNA 5′ end.

The HiScribe T7 Quick High Yield RNA Synthesis Kit (NEB #E2050) utilizes a master mix format, allowing for faster reaction setup. All that is required is the addition of two master mix reagents to your DNA template and water, reducing pipetting errors. DNase I and lithium chloride are included for DNA template removal and quick RNA purification.

For templates that contain an SP6 promoter, NEB offers the HiScribe SP6 RNA Synthesis Kit (NEB #E2070). This kit is designed for the in vitro transcription of RNA using SP6 RNA Polymerase. DNase I and lithium chloride are included for DNA template removal and quick RNA purification.

To determine the best kit for your application, please refer to the selection chart provided below.

T7 Kits SP6 Kits
Applications
Hiscribe T7 High Yield RNA Synthesis Kit (#E2040) Hiscribe T7 Quick Highyield RNA Synthesis Kit (#E2050) Hiscribe T7 ARCA mRNA Kit (#E2065) Hiscribe T7 ARCA mRNA (With Tailing) (#E2060) Hiscribe SP6 RNA Synthesis (#E2070)
Probe labeling
Fluorescent labeling: FAM, Cyanine (Cy) dyes, etc.
  • Fluorescent in situ hybridization (FISH)
 
   

Non-fluorescent labeling: Biotin, Digoxigenin

  • Insituhybridization
  • Blot hybridization with secondary detection
  • Microarray
 
   

High specific activity radiolabeling

  • Blot hybridization
  • RNase protection
     
mRNA & RNA for transfection

Streamlined mRNA synthesis with ARCA co-transcriptional capping and enzymatic poly(A) tailing

  • Transfection
  • Microinjection
  • In vitro translation
     

Streamlined ARCA capped RNA synthesis

  • Template encoded poly(A) tails
  • Non polyadenylated transcripts
  • Transfection
  • Microinjection
  • In vitro translation
   
 

Co-transcriptional capping with alternate cap analogs

  • Transfection
  • Microinjection
  • In vitro translation
 
   

Post-transcriptional capping with Vaccinia Capping System

  • Transfection
  • Microinjection
  • In vitro translation
   

Complete substitution of NTPs: 5-mC, pseudouridine, etc.

  • Induction of stem cell pluripotency
  • Modulation of cell fate or phenotype
  • Post translational capping with Vaccinia mRNA Capping System
     
Partial substitution of NTPs: 5-mC, pseudouridine, etc.  
Unmodified RNA  
   

Hairpins, short RNA, dsRNA

  • Gene knockdown
 
   
Structure, function, & binding studies

Complete substitution of NTPs

  • Aptamer selection
  • Isotopic labeling
     

Partial substitution of one or more NTPs

  • Aptamer selection
  • Structure determination
 
   

Unmodified RNA

  • SELEX
  • Structure determination
 
   

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