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Gentier Real-Time PCR System
quantification analysis interface.
3.3.4.3 Melting Curve
For intercalating dyes (such as SYBR Green) and non cleavable hybridization probes, the
fluorescence intensity is proportional to the amount of double-strand DNA (dsDNA). However,
these dsDNA including specific and non-specific PCR products, which means that the presence
of primer dimers and other non specific products can affect the quality of real time PCR data.
Melting curve show that the melting degree of dsDNA varies with the rising temperature.
Different dsDNA has different melting temperature T
m
(the temperature at which, 50% of the
DNA has dissociated from double stramded to single stranded) owing to different base
sequence, fragment length and GC content. Therefore, the melting curve is usually used to
analyze whether there are non-specific PCR products in PCR products.
The melting curve analysis interface consists of five functional modules:
Melting Curve, Melting
Peak, Sample Setting, Result Table
and
Heat Map
.
The melting curve analysis interface is
divided into four areas, the application software displays four functional modules (
Melting
Curve
,
Melting Peak
,
Sample Setting
,
Result Table
) by default, as shown in figure D 13.
