The RiboMAX™ Large Scale RNA Production System generates large quantities of capped or uncapped RNA transcripts suitable for in vitro or in vivo translation. RNA synthesized with RiboMAX™ Large Scale RNA Production System performs better for in vitro rabbit reticulocyte translation systems than RNA synthesized by standard methods. This is especially notable with high RNA concentrations that normally inhibit in vitro translation. With the reduction of components inhibitory to translation, there may be an advantage for other applications requiring biologically active RNA.
The RiboMAX™ Large Scale RNA Production System consistently produces about 10- to 20-fold more RNA than is produced with the standard Riboprobe® System. Because the RiboMAX™ Systems produce such large quantities of RNA, they are not recommended for generation of high-specific-activity RNA probes, due to the cost requirement of radiolabeled nucleotides.
The RiboMAX™ System reactions differ from those of the Riboprobe® Systems in three main ways: a HEPES (pH 7.5) buffer is used rather than a Tris-HCl (pH 7.9) buffer; rNTP and magnesium concentrations are elevated at levels appropriate for either SP6 or T7 RNA polymerase; and inorganic pyrophosphatase is included in the reaction.
Note: Use of the RiboMAX™ System for production of capped transcripts requires separate purchase of the Ribo m7G Cap Analog (Cat. P1711).
Applications of the RiboMAX™ Systems include:
- Generation of dsRNA and long RNAs for nonmammalian RNA interference studies.
- In vitro synthesis of tRNA and rRNA, and RNA virus genomes and ribozymes.
- Production of substrates for RNA splicing, studies of RNA secondary structure and RNA:protein interactions.
- Synthesis of antisense RNA
- Gurevich, V.V. et al. (1991) Anal. Biochem. 195, 207–13.
- Beckler, G.S. (1992) Promega Notes 39, 12–6.