A method for detecting point mutations or base substitutions in a nucleic acid polymer is especially useful for detecting such mutations in the highest melting domain (HMD) of a double-stranded nucleic acid polymer and is particularly suited for use with RNA. The method involves the steps of: (a) preparing a solution containing a double-stranded nucleic acid polymer comprising a duplex of a single-stranded nucleic acid polymer to be analyzed and a c The invention described and claimed in this application was made with government support under Grant #DK-38381 awarded by the National Institutes of Health. The U.S. government has certain rights in the invention.
The disclosure relates to a method for resolving double-stranded DNA species differing by at least one base pair. Each of the species is characterized by an iso-melting domain with a unique melting temperature contiguous with a melting domain of higher thermal stability.
Solvents for salt-gradient anion-exchange separation of nucleic acids, especially double-stranded DNA and especially by liquid chromatography, are improved by replacing NaCl as the eluting salt with any of a wide range of alkyl ammonium salts and can be used to elute nucleic acids in strict order of increasing length, with reduced sensitivity to elution temperature and salt concentration. Anion-exchange chromatography with these solvents is well suited for identification of DNA fragments on the basis of size, with greater accuracy, precision, and resolvable size range than often is possible with gel electrophoresis.
