What is Real-time PCR? Performing PCR in real time means that
product formation is measured at each cycle as it is occuring by
measuring a change in a fluorescent reporter signal. With
traditional, end-point PCR the signal is measured only once
after the cycling program has ended. Real-time PCR should not be
confused with RT-PCR, which is reverse transcription PCR, where
the starting template is RNA that is reverse transcribed into a
cDNA target prior to PCR. What kind of reporter can I use in my
real-time PCR assay? There are two general types of fluorescent
reporters that are used in real-time PCR assays: Double-stranded
DNA-binding Dyes Double-stranded DNA-binding dyes are dyes that
bind to the minor groove of double-stranded DNA (dsDNA). The
free dye has a very low fluorescence, whereas the bound dye has
a high fluorescence. The more double-stranded PCR product that
is produced, the greater the fluorescence signal will be. BRYT
Green® Dye is an example of a dsDNA-binding dye. These dyes
allow you to use s