T7-Scribe™ Standard RNA IVT Kit

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

The T7-Scribe™ Standard RNA IVT Kit is specially formulated to enable users to obtain the maximumpossible yields of canonical (GAUC) RNA from an in vitro transcription (IVT) reaction. The standard2 hour, 20 μl reaction will yield up to 150 μg of RNA from 1 μg of the control template. These yields aremade 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-Scribereaction can be readily modified to prepare fluorescent-, biotinylated- or digoxigenin-labeled RNA. T7-Scribe IVT RNA can be pr°Cessed into mRNA (5'-end capped and 3'-end poly(A) tailed) through theuse of CELLSCRIPT's ScriptCap™ m7G Capping System, ScriptCap 2'-O-Methyltransferase Kit andA-Plus™ Poly(A) Polymerase Tailing Kit (available separately). CELLSCRIPT also offers the INCOGNITO™ line of transcription kits for the production of pseudouridine-& 5-methyl-cytosine-containing (GA C and GA 5mC) IVT RNA. It has been shown that -mRNAs and5mC-mRNAs are translated into protein at higher levels and induce lower innate immune responses inhuman and other mammalian cells that express various RNA sensors compared t°Correspondingunmodified mRNAs

RNase-Free DNase I is provided in a 50% glycerol solution containing 50 mM Tris-HCl, pH 7.5, 10 mM CaCl2, 10 mM MgCl2 and 0.1% Triton® X-100. ScriptGuard RNase Inhibitor is supplied in a 50% glycerol solution containing 50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 10 mM DTT, 0.1 mM EDTA and 0.1% Triton X-100. All other enzymes are provided in a 50% glycerol solution containing 50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA and 0.1% Triton X-100.

Unit Definition One unit of RNase-Free DNase I digests 1 µg of pUC19 DNA to oligodeoxynucleotides in 10 minutesat 37°C. One unit of ScriptGuard RNase Inhibitor results in 50% inhibition of 5 ng of RNase A. Activity ismeasured by the inhibition of hydrolysis of cyclic 2',3'-CMP by RNase A. Functional Testing The T7-Scribe Standard RNA IVT Kit is functionally tested under standard reaction conditions usingthe T7 Control Template DNA. The kit must produce at least 136 µg of RNA from 1 µg of the T7Control Template DNA in 2 hours at 37°C. Contaminating Activity Assays All components of the T7-Scribe Standard RNA IVT Kit are free of detectable RNase and DNaseactivity, except for the inherent activity of the RNase-Free DNase I component.