Procedure miniaturized in multiwell plates to analyze the binding of ligands to nucleic acids by obtaining thermal melting curves.

摘要

Procedure miniaturized in multiwell plates to analyze the binding of ligands to nucleic acids by obtaining thermal melting curves.; a method miniaturized in multiwell plates to analyze the binding of ligands to nucleic acids by thermal melting curves described. To achieve miniaturization fluorescent labeling strategies that allow the determination of the proportion of associated complex and dissociated in homogeneous phase was used. The reaction mixtures to be analyzed are placed in individual wells of a multiwell plate. The plate is subjected to a gradual increase in temperature programmed time, taking readings of fluorescence intensity of each well after each step of temperature increase. It will also run a program of gradual temperature decrease, also taking readings of fluorescence intensity with each change. The programmed temperature change and reading fluorescence emission intensity can be performed in an apparatus for real-time PCR or other equipment to perform such operations on multiwell plates.