AmpliCap™ SP6 High Yield Message Maker Kit

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

The AmpliCap™ SP6 High Yield Message Maker Kit produces capped RNA by in vitro transcription using SP6 RNA polymerase and the standard cap analog m7G[5']ppp[5']G. A 2 hour, 20 μl AmpliCap reaction yields up to 35 μg of m7G-capped RNA from 1 μg of the 1.4 kb standard SP6 control template DNA. The m7G[5']ppp[5']G cap analog/NTP PreMix contains all four ribonucleotides and the monomethylated cap analog. The PreMix ensures the optimal concentration of each NTP and ratio of methylated cap analog to GTP (4:1), maximizing transcript capping (~80%) and yield. Because the concentration of GTP in the reaction is limiting, the cap analog is preferentially incorporated as the first or 5'-terminal G of the transcript. CELLSRIPT also offers the AmpliCap-Max™ T7 High Yield Message Maker Kit and the MessageMAX™ T7 ARCA-Capped Message Transcription Kit, both of which produce unsurpassed yields of capped in vitro transcripts (up to 60 μg of RNA in a 30 minute standard control reaction with an 80% capping efficiency). The AmpliCap-Max Kit uses the standard RNA Cap Analog m7G[5']ppp[5']G and the MessageMAX Kit uses the anti-reverse cap analog m27,3'-OG[5']ppp[5'].

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 a50% glycerol solution containing 50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 10 mM DTT, 0.1 mM EDTAand 0.1% Triton® X-100. All other enzymes are provided in a 50% glycerol solution containing 50 mMTris-HCl, pH 7.5, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA and 0.1% Triton X-100.

Unit Definitions 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 AmpliCap SP6 High Yield Message Maker Kit is functionally tested under standard reactionconditions using the SP6 Control Template DNA. The kit must produce at least 30 µg of RNA from1 µg of the SP6 Control Template DNA in 2 hours at 37°C. Contaminating Activity Assays All components of the AmpliCap SP6 High Yield Message Maker Kit are free of detectable RNase andDNase activity, except for the inherent activity of the RNase-Free DNase I component.

http://www.cellscript.com/products/019pl0114CS.pdf