Applications

In vitro Transcription: In Vitro Transcription

ScriptCap™ m7G Capping System

For post-transcriptional capping of in vitro transcribed RNA with ScriptCap Capping Enzyme, GTP and SAM. Cap 0 (N7-methyl-G) caps can be built with efficiencies approaching 100%. This cannot be accomplished with co-transcriptional capping methods. Use in conjunction with ScriptCap 2'-O-Methyltransferase to produce Cap 1 caps, that which is found on most eukaryotic mRNAs.

CellScript

Catalogue No.	Description	Pack Size	Price	Qty	Change to: €
C-SCCE0625	ScriptCap™ m⁷G Capping System	25 Reactions	£450.00	Quantity	Add to Order

Description

Achieve ~100% Cap 0 capping of IVT RNA, a level unattainable using cap analogs.
Enhance mRNA stability and translation efficiency through post-transcriptional 5’ capping.
Use simultaneously with ScriptCap™ 2’-O-Methyltransferase to produce Cap 1 caps in a single reaction.
Simplified workflow with direct input of capped RNA into A-Plus™ Poly(A) Polymerase tailing reactions with no cleanup required.

Product is for research use only (RUO)

Product Description:

The ScriptCap™ m⁷G Capping System adds a methylated guanine nucleotide cap to the 5′ end of RNA, generating Cap 0-RNA with ~100% efficiency. The process consists of three enzymatic reactions: (1) conversion of the 5′ triphosphate of RNA to a diphosphate, (2) joining of GTP to the 5′ diphosphate of the first nucleotide and (3) methylation of the 7-nitrogen of guanine using S-adenosyl-methionine (SAM). These reactions produce Cap 0-RNA with nearly 100% efficiency, a level that cannot be obtained using co-transcriptional capping methods. A standard reaction caps approximately 60 μg of RNA and can be scaled up or down to accommodate user needs.

5’ Capping enhances mRNA stability and translation efficiency in cells compared to uncapped RNA. Cap 0-RNA can be converted to Cap 1-RNA using the ScriptCap™ 2′-O-Methyltransferase Kit to further boost in vivo translation efficiency. ScriptCap™ capped RNA can be added directly to A-Plus™ Poly(A) Polymerase tailing reactions without cleanup for seamless synthesis of fully 5’ capped and 3’ tailed mRNA.

Materials Supplied:

Important Store at -20°C in a freezer without a defrost cycle. Do not store at -70°C.

ScriptCap™ m⁷G Capping System Contents (25 reactions)
Kit Component	Reagent Volume
ScriptCap™ Capping Enzyme, 10 U/μl in 50% glycerol, 50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1 mM dithiothreitol (DTT), 0.1 mM EDTA and 0.1% Triton® X-100	100 μl
10X ScriptCap™ Capping Buffer 0.5 M Tris-HCl, pH 8.0, 60 mM KCl and 12.5 mM MgCl₂	250 μl
10 mM GTP	250 μl
20 mM S-adenosyl-methionine (SAM)	30 μl
ScriptGuard™ RNase Inhibitor, 40 U/μl in 50% glycerol, 50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 10 mM DTT, 0.1 mM EDTA and 0.1% Triton X-100	65 μl
RNase-Free Water	2 x 1.4 ml

Materials Required, but not Supplied

IVT RNA
Materials or kits for purification of the RNA product