Novel transcription reaction conditions and Ambion's patented, high-yield technology allow the synthesis of large mass amounts of RNA produced from small templates (<300 bases). Each Ambion® kit includes sufficient reagents for 25 reactions.
• Efficiently transcribes small templates (<300 bases) with consistently high yields
• Designed for use with plasmid, synthetic oligonucleotide, or PCR product templates
Short transcripts require many more transcription initiation events to synthesize a given mass of product than do longer transcripts. Since the initiation step is the rate-limiting step in transcription reactions, the yield of product from short templates is often low. The MEGAshortscript™ High Yield Transcription Kit has been designed to maximize the yield of short RNA transcripts from plasmid, oligonucleotide, and PCR templates. MEGAshortscript™ Kits are only optimized for use with the T7 polymerase included in the kit. Using a different polymerase other than that which is supplied in the kit may result in low yields.
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![Yields of MEGAsho...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/f00187.jpg","Yields of MEGAshortscript™ vs Large Scale MAXIscript™ Reactions - Panel A","Yields of a 100-base transcript with MEGAshortscript and MAXIscript (large scale synthesis conditions) in vitro transcription kits. The template was a 125 bp PCR product containing a T7 promotor. Reactions were carried out with a trace amount of [32P] UTP at 37°C. 1 µl aliquots of each reaction were removed after 2 hours and 4 hours, and the percent incorporation of [32P] UTP assessed by TCA precipitation to determine the yield. Transcript size and integrity of the reactions were assessed by electrophoresis of 1 µl aliquots of each 2 hr reaction on a 10% polyacrylamide/8 M urea gel. The specific activity of all transcripts was 1 x 105 cpm/µg. Lanes 1-5: MEGAshortscript reactions containing increasing concentrations of template DNA: 50 nM (41 ng); 100 nm (83 ng); 200 nM (166 ng); 300 nM (249 ng); 400 nM (332 ng). Lanes 6-10: MAXIscript reactions containing the same increasing amounts of template DNA. Note: at low concentrations, template is limiting for MEGAshortscript reactions.");)
![Yields of MEGAsho...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/f00188.jpg","Yields of MEGAshortscript™ vs Large Scale MAXIscript™ Reactions - Panel B","Yields of a 100-base transcript with MEGAshortscript and MAXIscript (large scale synthesis conditions) in vitro transcription kits. The template was a 125 bp PCR product containing a T7 promotor. Reactions were carried out with a trace amount of [32P] UTP at 37°C. 1 µl aliquots of each reaction were removed after 2 hours and 4 hours, and the percent incorporation of [32P] UTP assessed by TCA precipitation to determine the yield. Transcript size and integrity of the reactions were assessed by electrophoresis of 1 µl aliquots of each 2 hr reaction on 10% polyacrylamide/8 M urea gel. The specific activity of all transcripts was 1 x 105 cpm/µg. This graph shows the yields for reactions 1-10 in the previoue figure after 2 hours and 4 hours.");)
