Vaccinia Capping Enzyme

This capping enzyme contains three enzymatic activities (RNA triphosphatase, guanylyltransferase and guanine methyltransferase) to forming a Cap 0 structure (m7Gppp5'N) In the presence of the vaccinia capping enzyme, GTP and SAM (methyl donor) the In Vitro Transcripts mRNA can be efficient add capped in less one hour.
No. Size Price Qty Status
C15037-500U set $240.00 Inquiry
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Escherichia coli
>95% as determined by SDS-PAGE. Purified by Ni-NTA chromatography.
Unit Definition:
One unit of Vaccinia Capping Enzyme is defined as the amount of enzyme required to incorporate 10 pmol of (α32P) GTP
into an 80 nt transcript in 1 hour at 37°C.
Reaction Condition:
1X Capping enzyme reaction buffer, supplemented with 0.5 mM GTP and 0.1 mM S-adenosylmethione (SAM). Incubate at 37°C.
10X Capping enzyme Reaction Buffer: 500 mM Tris-HCl (pH 8.0), 50 mM KCl, 10 mM MgCl2, 10 mM DTT.
Storage Buffer:
Capping enzyme is supplied in 20 mM Tris-HCl (pH 8.0), 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.1% Triton® X-100 and 50% Glycerol.

Stored at -20°C. Avoid repeated freeze/thaw cycles..

Reaction should be performed under RNase free condition. Use nuclease-free tubes, reagents and water to avoid RNase contamination. Also, wear gloves when working with RNA.
Capping procedures:
1. Combine RNA and nuclease-free H2O to a final 15 μL.
2. Heating at 65°C for 5 minutes then chill on ice for 5 minutes.
3. Below reaction mixture should be prepared on ice and combined in the following order:
Component Amount Final concentration
Denatured RNA 15 μL -
10X Capping Buffer 2 μL 1X
10 mM GTP 1 μL 0.5 mM
SAM (2 mM) 1 μL 0.1 mM
Vaccinia Capping Enzyme 1 μL 10 U/rxn
4. Gently mix the reaction thoroughly to achieve uniform distribution.
5. Incubate at 37°C for 30 minutes.