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RiboMAX™ Large Scale RNA Production Systems

RiboMAX™ Large Scale RNA Production Systems

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Description

Size

Catalog number : P1280

Catalog number :  P1300

More RNA from Transcription Reactions

The RiboMAX™ Large Scale RNA Production System synthesizes large quantities of capped or uncapped RNA transcripts suitable for in vitro or in vivo translation. RNA synthesized with the 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.

RiboMAX™ System reactions differ from those of the Riboprobe® Systems in three main ways: 1) a HEPES (pH 7.5) buffer is used rather than a Tris-HCl (pH 7.9) buffer; 2) rNTP and magnesium concentrations are elevated at levels appropriate for either SP6 or T7 RNA polymerase; and 3) 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).

RiboMAX™ System Applications:

  • 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

References

  1. Gurevich, V.V. et al. (1991) Anal. Biochem. 195, 207–13.
  2. Beckler, G.S. (1992) Promega Notes 39, 12–6.
RiboMax produces more RNA than standard in vitro transcription protocols.
Effect of pGEM®-luc RNA concentration on in vitro translation. The indicated amounts of pGEM®-luc RNA transcripts were used to program 50µl of Rabbit Reticulocyte Lysate in vitro translation reactions. Expression of functional luciferase was measured as follows: 2.5µl of each reaction was mixed with an equal volume of Luciferase Assay Reagent and luminescence measured for 10 seconds in a luminometer.

Protocols

Complete Protocol

RiboMAX™ Large Scale RNA Production Systems—SP6 and T7 Technical Bulletin


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Quick Protocol

RiboMAX Large Scale RNA Production Systems Quick Protocol FB059


Specifications

What’s in the box?

Item / Catalog number: P1280 Part # Size Concentration Available Separately
rATP, 100mM E601D 1 × 100μl 100mM View Product
rUTP, 100mM E602D 1 × 100μl 100mM View Product
rGTP, 100mM E603D 1 × 100μl 100mM View Product
rCTP, 100mM E604D 1 × 100μl 100mM View Product
SP6 Transcription Buffer, 5X P131A 1 × 240μl
SP6 Linear Control DNA P134A 1 × 10μl
Sodium Acetate, 3.0M (pH 5.2) P135A 1 × 1ml
Enzyme Mix, SP6 RNA Polymerase P133A 1 × 120μl
Nuclease-Free Water P119A 1 × 1,250μl View Product
RQ1 RNase-Free DNase M610C 1 × 110u View Product

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

 

What’s in the box?

Item / Catalog number: P1300 Part # Size Concentration Available Separately
RQ1 RNase-Free DNase M610C 1 × 110u View Product
rATP, 100mM E601D 1 × 100μl 100mM View Product
rUTP, 100mM E602D 1 × 100μl 100mM View Product
rGTP, 100mM E603D 1 × 100μl 100mM View Product
rCTP, 100mM E604D 1 × 100μl 100mM View Product
Nuclease-Free Water P119A 1 × 1,250μl View Product
Sodium Acetate, 3.0M (pH 5.2) P135A 1 × 1ml
Enzyme Mix, T7 RNA Polymerase P137A 1 × 120μl
T7 Transcription Buffer, 5X P140A 1 × 240μl 5X
T7 Linear Control DNA P141A 1 × 10μl

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB