Methods are provided for nucleic acid analysis wherein a target nucleic
acid that is at least partially double stranded is mixed with a dsDNA
binding dye having a percent saturation of at least 50% to form a
mixture. In one embodiment, the nucleic acid is amplified in the presence
of the dsDNA binding dye, and in another embodiment a melting curve is
generated for the target nucleic acid by measuring fluorescence from the
dsDNA binding dye as the mixture is heated. Dyes for use in nucleic acid
analysis and methods for making dyes are also provided.