69330  Red Nova Lysate Control RNA

Flexibility for optimization of ion concentrations

Red Nova® Lysate is a complete kit designed for efficient cell-free synthesis of proteins from exogenous RNA templates. Prepared from rabbit reticulocytes, the lysate is treated with a calcium-dependent nuclease to remove endogenous mRNA. Special care has been taken to optimize this treatment to maximize translation activity with added templates from natural and synthetic sources. The Red Nova Lysate Kit contains all necessary reaction components; simply provide the test RNA and the appropriate radioactive amino acid.

Because the translation efficiency and fidelity of initiation of particular RNAs depend on the salt concentrations and counter ions used in the reaction, magnesium and potassium solutions are provided separately. The system includes KCl as the salt of choice for pCITE^® transcripts and higher fidelity translation of capped mRNAs and KOAc for improved performance of uncapped messages (Jackson 1991). A unique Control RNA is included, which permits monitoring of translation performance and yields a set of 6 protein size markers when translated in vitro. One domain of the initial translation product is a specific protease that cleaves the rest of the molecule into discrete peptides during the translation reaction. Their final sizes are 6.3, 21.8, 30.3, 52.0, 73.7, and 83.3 kDa, which are convenient sizes for accurate gel analysis of experimental samples.