Glossary
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
220
melt curve
A plot of data collected during the melt curve stage. Peaks in the melt curve can indicate the
melting temperature (Tm) of the target or can identify nonspecific PCR amplification. In
the 7500 software, you can view the melt curve as normalized reporter (Rn) vs. temperature
or as derivative reporter (
−
Rn
′
) vs. temperature. Also called dissociation curve.
melt curve stage
In the thermal profile, a stage with a temperature increment to generate a melt curve.
melting temperature
(Tm)
In melt curve experiments, the temperature at which 50% of the DNA is double-stranded
and 50% of the DNA is dissociated into single-stranded DNA. The Tm is displayed in the
melt curve.
multicomponent
plot
A plot of the complete spectral contribution of each dye for the selected well(s) over the
duration of the PCR run.
negative control
(NC)
In the 7500 software, the task for targets or SNP assays in wells that contain water or buffer
instead of sample. No amplification of the target should occur in negative control wells.
Previously called no template control (NTC).
negative control-
blocked IPC wells
In presence/absence experiments, wells that contain IPC blocking agent instead of sample
in the PCR reaction. No amplification should occur in negative control-blocked IPC wells
because the reaction contains no sample and amplification of the IPC is blocked. Previously
called no amplification control (NAC).
negative control-
IPC wells
In presence/absence experiments, wells that contain IPC template and buffer or water
instead of sample. Only the IPC template should amplify in negative control-IPC wells
because the reaction contains no sample. Previously called IPC+.
no amplification
control (NAC)
See negative control-blocked IPC wells.
no template control
(NTC)
See negative control (NC).
nonfluorescent
quencher-minor
groove binder (NFQ-
MGB)
Molecules that are attached to the 3
′
end of TaqMan
®
probes. When the probe is intact, the
nonfluorescent quencher (NFQ) prevents the reporter dye from emitting fluorescence.
Because the NFQ does not fluoresce, it produces lower background signals, resulting in
improved precision in quantitation. The minor groove binder (MGB) increases the melting
temperature (Tm) without increasing probe length. It also allows the design of shorter
probes.
normalized quantity
Quantity of target divided by the quantity of endogenous control.
normalized reporter
(Rn)
Fluorescence from the reporter dye normalized to the fluorescence of the passive reference.
omit well
An action that you perform before reanalysis to omit one or more wells from analysis.
Because no algorithms are applied to omitted wells, omitted wells contain no results.
outlier
For a set of data, a datapoint that is significantly smaller or larger than the others.
