ABSTRACT
Upon heating, double-stranded DNA (dsDNA) melts into single-stranded random coils. The DNA unfolding, which occurs within a narrow temperature range, can be monitored in real time by use of generic uorescent dyes which have high afnity to dsDNA.1 These specialized dyes have low uorescence when they are free in solution and a large uorescence emission upon binding to dsDNA.2 A characteristic DNA melting curve can be generated by plotting the decrease in uorescence as the temperature of the sample is increased, such that the duplex dissociation temperature is reached and the dye is released (Figure 28.1a).
