From the software, you can use the plate layout as a selection tool to assign well
contents, to view well assignments, and to view results. The plate layout can be
printed, included in a report, exported, and saved as a slide for a presentation.
One standard in a standard curve. The standard quantity for each point in a standard
curve is calculated based on the starting quantity and serial factor.
In genotyping and presence/absence experiments, a DNA sample with a known
genotype, homozygous or heterozygous.
In the software, the task for the SNP assay in wells that contain a sample with a
known genotype.
Fluorescent compound used to calibrate the instrument. See “system dye“ on
In quantification experiments, the amount of target in the samples. Absolute quantity
can refer to copy number, mass, molarity, or viral load. Relative quantity refers to the
fold-difference between normalized quantity of target in the sample and normalized
quantity of target in the reference sample.
A molecule attached to the 3′ end of probes to prevent the reporter from emitting
fluorescence signal while the probe is intact. With reagents, a nonfluorescent
quencher-minor groove binder (NFQ-MGB) can be used as the quencher.
Regression coefficient calculated from the regression line in the standard curve. An
important quality value, the R
2
value indicates the closeness of fit between the
standard curve regression line and the individual C
t
data points from the standard
reactions. A value of 1.00 indicates a perfect fit between the regression line and the
data points.
A plot of raw fluorescent signal as detected through each emission filter, used to view
raw data for individual wells and at individual cycles.
A solution that contains all components to run the PCR reaction, except for the
template (sample, standard, or control). Also called a “PCR cocktail”.
The PCR reaction components used to amplify the target and to detect amplification.
Process of collecting fluorescence data during PCR. Data from the real-time PCR are
used to calculate results for quantification experiments or to troubleshoot results for
genotyping or presence/absence experiments.
In relative standard curve and Comparative C
t
(ΔΔC
t
) experiments, the sample used
as the basis for relative quantification results. Also called the calibrator.
Type of calibration in which the software maps the positions of the wells on the
sample block of the instrument. The software uses the ROI calibration data to
associate increases in fluorescence during a run with specific wells of the plate or
array card. A calibration image for each individual filter must be generated to account
for minor differences in the optical path.
point
positive control
pure dye
quantity
quencher
R
2
value
Raw Data Plot
reaction mix
reagents
real-time PCR
reference sample
region of interest
(ROI) calibration
Limited product warranty
QuantStudio
™
6 and 7 Flex Real-Time PCR Systems v1.6 Maintenance and Administration Guide
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