RiboMAX Large Scale RNA Production System-SP6

More RNA from Transcription Reactions

The RiboMAX™ Large Scale RNA Production Systems consistently produce 2–5mg/ml of RNA in a 1ml reaction, about 10- to 20-fold more RNA than is produced with the standard Riboprobe® System transcription reaction. 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.

RNAs synthesized with the RiboMAX™ System perform better for in vitro translation in rabbit reticulocyte translation systems than RNA synthesized by standard methods. The reduction of components inhibitory to translation may be an advantage for other applications requiring biologically active RNA. Because the RiboMAX™ Systems produce large quantities of RNA, they are not recommended for generating high-specific-activity RNA probes.

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 large quantities of capped or uncapped RNA transcripts for in vitro or in vivo translation.
• In vitro synthesis of tRNA, rRNA, RNA virus genomes and ribozymes.
• Production of substrates for RNA splicing, RNA secondary structure, antisense RNA and RNA:protein interaction studies.
• Synthesis of long RNAs for nonmammalian RNA interference.

References

1. Gurevich, V.V. et al. (1991) Anal. Biochem. 195, 207–13.
2. Beckler, G.S. (1992) Promega Notes 39, 12–6.