HiScribe™ T7 Quick High Yield RNA Synthesis Kit
- Up to 180 μg of RNA generated per reaction, from 1 μg of control template
- Master mix format reduces pipetting steps while still enabling partial substitution of NTPs for labeling and incorporation of modified bases
- Template removal and RNA purification reagents included
- Linearized control template included for verification of RNA synthesis
In Vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection
The HiScribe T7 Quick High Yield RNA Synthesis Kit is designed for quick set-up and production of large amounts of RNA in vitro. The reaction can be set up conveniently by combining the NTP buffer mix, T7 RNA Polymerase mix and a suitable DNA template. The kit also allows for capped RNA or dye-labeled RNA synthesis by incorporation of cap analog (ARCA, NEB #S1411) or dye-modified nucleotides. RNA synthesized with the kit can be used for RNA structure and function studies, ribozyme biochemistry, as probes for RNase protection assays and hybridization based blots, anti-sense RNA and RNAi experiments, microarray analysis and microinjection, as well as in vitro translation and RNA vaccines.
To synthesize high specific activity radioactive RNA probes or RNA with 100% substitution of one or more modified nucleotides we recommend using the T7 High Yield RNA synthesis kit (NEB #E2040), in which the four nucleotides are supplied separately.
DNA Template Preparation:
Linearized plasmid DNA, PCR products or synthetic DNA oligonucleotides can be used as templates for in vitro transcription with the HiScribe T7 Quick High Yield RNA Synthesis Kit, provided that they contain a double-stranded T7 promoter region upstream of the sequence to be transcribed. Figure 1 illustrates the minimal T7 promoter sequence, as well as a run-off transcript after T7 transcription.
The following reagents are supplied with this product:
Store at (°C) Concentration LiCl Solution -20 T7 RNA Polymerase Mix -20 DNase I (RNase-free) -20 2 units/μl FLuc Control Template 0.5 μg/μl NTP Buffer Mix -20 20 mM of each NTP
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