Using the MEGAscript Kit
, you can produce over 120 µg of RNA from 1 µg of DNA template. This is 10-50 times the amount of RNA produced by conventional transcription (Figure 2). Reactions are versatile and can be manipulated to include specialized reagents such as modified nucleotides or additional polymerase. In addition, Ambion has optimized the transcription of short (20-500 nt) DNA templates with the MEGAshortscript™ Kit
. This patented technology was designed to maximize the yield of short RNA transcripts from plasmid, oligonucleotides, and PCR product templates.
Electropherogram of RNA Transcripts of Different Sizes. In vitro transcription was performed using 1, 4, or 9 kb templates with Ambion's MEGAscript® Kit according to protocol. 20 µl reactions were diluted to 100 µl and 1 µl was assessed on the Agilent 2100 bioanalyzer. The single primary peak is indicative of product quality while the area under the peak is related to yield.
MEGAscript® vs. Conventional In Vitro Transcription Reactions. 20 µl transcription reactions containing 1 µg template, T7 MEGAscript components and [32
P]UTP, were incubated at 37°C. 1 µl aliquots were removed at 1 hr intervals and acid precipitable cpm determined by scintillation counting.