AI Article Synopsis
- The study investigated how Mg2+ facilitates RNA degradation during reverse transcriptase reactions using agarose gel electrophoresis to monitor the process.
- Both natural ribosomal poly(A)+ RNA and synthetic poly(rA) are similarly susceptible to degradation, indicating the reaction is not reliant on any specific RNA structure.
- The degradation rate increases with higher Mg2+ concentrations and is significantly enhanced by raising the pH, which offers insights for optimizing reverse transcriptase conditions.
Article Abstract
RNA degradation catalyzed by Mg2+ was studied under the conditions of reverse transcriptase reaction. Agarose gel electrophoresis in 6 M urea was employed to follow the reaction. Natural ribosomal of poly(A)+ RNA as well as synthetic poly(rA) are equally accessible for degradation. Neither an RNAase nor alkali alone is responsible for the degradation. The reaction rate is directly proportional to Mg2+ concentration in the range of 1 to 10 mM, doesn't depend upon RNA concentration and enhances approximately 50 fold upon increasing of pH value from 7.5 to 9.0. It was concluded that the Mg2+ catalyzed degradation reaction is an unspecific one in respect to the primary structure of RNA. The results obtained are useful to optimize the conditions for the reverse transcriptase reaction.
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