The SP6-Scribe™ Standard RNA IVT Kit is specially formulated to enable users to obtain the maximum possible yields of canonical (GAUC) RNA from an in vitro transcription (IVT) reaction. The standard 2 hour, 20 μl reaction will yield up to 90 μg of RNA from 1 μg of the control template. These yields are made possible by the high-performance properties of the SP6-Scribe enzyme.

 The SP6-Scribe Standard RNA IVT Kit produces exceptionally high yields of either long or short transcripts. The standard reaction can be scaled up to produce milligram amounts of RNA. In addition, an SP6-Scribe reaction can be readily modified to prepare fluorescent-, biotinylated- or digoxigenin-labelled RNA.

SP6-Scribe IVT RNA can be processed into mRNA (5'-end capped and 3'-end poly(A) tailed) through the use of CELLSCRIPT's ScriptCap™ m7G Capping System, ScriptCap 2'-O-Methyltransferase Kit and A-Plus™ Poly(A) Polymerase Tailing Kit (available separately).

The T7-Scribe™ Standard RNA IVT Kit is specially formulated to enable users to obtain the maximum possible yields of canonical (GAUC) RNA from an in vitro transcription (IVT) reaction. The standard 2 hour, 20 μl reaction will yield up to 150 μg of RNA from 1 μg of the control template. These yields are made possible by the high-performance properties of the T7-Scribe enzyme.

The T7-Scribe Standard RNA IVT Kit produces exceptionally high yields of either long or short transcripts. The standard reaction can be scaled up to produce milligram amounts of RNA. In addition, a T7 -Scribe reaction can be readily modified to prepare fluorescent-, biotinylated- or digoxigenin-labelled RNA.

T7-Scribe IVT RNA can be processed into mRNA (5'-end capped and 3'-end poly(A) tailed) through the use of CELLSCRIPT's ScriptCap™ m7G Capping System, ScriptCap 2'-O-Methyltransferase Kit and A-Plus™ Poly(A) Polymerase Tailing Kit (available separately).

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Protocols for: SP6-Scribe™ and T7-Scribe™ Standard RNA IVT kits

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1.Karikó, K. et al., (2008) Molecular Therapy 16, 1833.

2.Anderson, B.R. et al., (2010) Nucl. Acids Res. 17, 5884.

3.Karikó, K. et al., (2005) Immunity 23, 165.

4.Karikó, K. and Weissman, D. (2007) Curr. Opin. Drug Discov. Devel. 10, 523.

5.Sambrook, J. et al., (1989) Molecular Cloning: A Laboratory Manual (2nd ed.), New York, Cold Spring Harbor Laboratory Press

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• Use AmpliCap™ High Yield Message Maker Kits for efficient production of RNA with an m⁷G[5´]ppp[5´]G cap structure on the 5´ end.
• Because of the very high concentration of NTPs used in AmpliScribe reactions, these kits are not recommended for preparing high specific-activity radiolabeled RNA probes.

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Applications

• High-yield synthesis of RNA from DNA cloned downstream from a T7, T3, or SP6 RNA polymerase promoter, for microinjection, RNA interference, or antisense experiments.
• Structural and functional studies of splicing, processing, heterogeneous nuclear RNAs, tRNAs, viral RNAs, and ribozymes.¹
• Production of fluorescent²-, biotinylated³-, or digoxigenin⁴- labeled RNA

Benefits

• High yields of RNA from all templates—synthesize both long (Fig. 2) and short RNA using only one kit.⁵ Produce microgram amounts of RNA from as little as 1 ng of template.
• Reactions can be scaled up to produce milligram amounts of RNA.
• Produce >20 times more RNA than conventional or "homemade" in vitro transcription reactions.
• RNase inhibitor included.

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