The HiScribe T7 Quick High Yield RNA Synthesis Kit is designed for quick set-up andproduction of large amounts of RNA in vitro. The reaction can be set upconveniently by combining the NTP buffer mix, T7 RNA Polymerase mix and asuitable DNA template. The kit also allows for capped RNA or dye-labeled RNAsynthesis by incorporation of cap analog (ARCA, NEB #S1411) or dye-modifiednucleotides. RNA synthesized with the kit can be used for RNA structureand function studies, ribozyme biochemistry, as probes for RNase protectionassays and hybridization based blots, anti-sense RNA and RNAi experiments,microarray analysis and microinjection, as well as in vitro translation and RNAvaccines.To synthesize high specific activity radioactive RNA probes or RNA with 100%substitution of one or more modified nucleotides we recommend using theT7 High Yield RNA Synthesis Kit (NEB #E2040), in which the four nucleotides aresupplied separately.DNA Template Preparation:Linearized plasmid DNA, PCR products or synthetic DNA oligonucleotides canbe used as templates for in vitro transcription with the HiScribe T7 Quick High YieldRNA Synthesis Kit, provided that they contain a double-stranded T7 promoterregion upstream of the sequence to be transcribed. Figure 1 illustrates theminimal T7 promoter sequence, as well as a run-off transcript after T7 transcription.Figure 1. Transcription by T7 RNA Polymerase
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