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Translation Optimization for Mg2+ and K+ Concentrations
The Flexi® Rabbit Reticulocyte Lysate System allows translation reactions to be optimized for a wide range of parameters, including Mg2+ and K+ concentrations and the choice to add or omit DTT. To help optimize Mg2+ for a specific message, the endogenous Mg2+ concentration of each lysate batch is stated in the product information included with this product. The Flexi® System also offers the choice of three amino acid mixtures and includes a control RNA encoding the firefly luciferase gene.
The Flexi® Rabbit Reticulocyte Lysate System, when optimized, can increase the quantity of protein produced as much as fourfold over a standard lysate reaction. The Flexi® System contains three Amino Acid Mixtures, enabling different choices of radioisotopes. Luciferase Control RNA and Luciferase Assay Reagent are included with the system as a functional control (only full-length luciferase is active).
For more information, see the Protocols & Applications Guide.
Applications:
- Optimizing translation of low-expressing messages.
- Oxidizing environment can be adjusted, which may be important for maintaining functional activity related to disulfide bridges.
- Luciferase activity can be used for the detection of inhibitory contaminants in RNA preparations.
- Investigation of translational control.
- Screening of chemical libraries for effect on translation.
Reference:
- Beckler, G.S. et al. (1995) Methods in Molecular Biology 37, 215–32.
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Protocols
Complete Protocol
Flexi® Rabbit Reticulocyte Lysate Systems Technical Bulletin
PDF (1 MB)
Quick Protocols
Flexi Rabbit Reticulocyte Lysate System Quick Protocol FB041
PDF (91 KB)
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Certificate of Analysis
Lookup Certificate of AnalysisStorage Conditions
LESS THAN -65C
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.
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Resources
Citations
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Selection of mRNA 5´-untranslated region sequence with high translation efficiency through ribosome display.
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2008 Biochem. Biophys. Res. Commun.
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Selection of mRNA 5´-untranslated region sequence with high translation efficiency through ribosome display.