Nucleic acid separation and detection by electrophoresis with a counter-migrating high-affinity intercalating dye

摘要

The invention provides novel electrophoretic methods for high-resolution separation and high-sensitivity detection of nucleic acids. The methods involve the use of a high-resolution buffer and a counter-migrating high-affinity intercalating dye in an electrophoresis system to achieve superior separation and detection of nucleic acids. A kit for separation and detection of nucleic acids in a sample by electrophoresis is also provided. The kit comprises a gel buffer containing a high-resolution intercalating dye with a resolution of at least 2.0 between the 271-bp and 282-bp phiX 174 HaeIII nucleic acid fragments nucleic acid fragments; and a high-affinity intercalating dye having a positive charge and DNA affinity constant of at least 10<6>M<-1>.