Glossary
219
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
experiment name
Entered during experiment setup, the name that is used to identify the experiment.
Experiment names cannot exceed 100 characters and cannot include any of the following
characters: forward slash (/), backslash (\), greater than sign (>), less than sign (<), asterisk
(*), question mark (?), quotation mark ("), vertical line (|), colon (:), or semicolon (;).
experiment type
The type of experiment you are performing using the 7500/7500 Fast system:
• Standard curve
• Comparative C
T
(
∆∆
C
T
)
• Relative standard curve
• Melt curve (not available in the Design Wizard)
• Genotyping
• Presence/absence
The experiment type you select affects the setup, run, and analysis.
forward primer
Oligonucleotide that flanks the 5
′
end of the amplicon. The reverse primer and the forward
primer are used together in PCR reactions to amplify the target.
holding stage
In the thermal profile, a stage that includes one or more steps. You can add a holding stage
to the thermal profile to activate enzymes, to inactivate enzymes, or to incubate a reaction.
housekeeping gene
A gene that is involved in basic cellular functions and is constitutively expressed.
Housekeeping genes can be used as endogenous controls. See also endogenous control.
internal positive
control (IPC)
In presence/absence experiments, a short synthetic DNA template that is added to PCR
reactions. You can use the IPC to distinguish between true negative results (that is, the target
is absent in the samples) and negative results caused by PCR inhibitors, incorrect assay
setup, or reagent or instrument failure.
inventoried assays
TaqMan
®
Gene Expression Assays and TaqMan
®
SNP Genotyping Assays that have been
previously manufactured, passed quality control specifications, and stored in inventory.
IPC
In presence/absence experiments, abbreviation for internal positive control (IPC). In the
7500 software, the task for the IPC target in wells that contain the IPC and do not contain
IPC blocking agent. See also internal positive control (IPC).
IPC blocking agent
Reagent added to PCR reactions to block amplification of the internal positive control
(IPC).
IPC+
See negative control-IPC wells.
made-to-order
assays
TaqMan
®
Gene Expression Assays or TaqMan
®
SNP Genotyping Assays that are
manufactured at the time of order. Only assays that pass manufacturing quality control
specifications are shipped.
manual baseline
An analysis setting in which you enter the baseline start and end values for the amplification
plot. You can apply the manual baseline setting to specific wells in the reaction plate.
manual C
T
An analysis setting in which you enter the threshold value and select whether to use
automatic baseline or manual baseline values. The software uses the baseline and the
threshold values to calculate the threshold cycle (C
T
).
