DRAFT
September 25, 2007 1:07 am, 4376782_Glossary.fm
Glossary
139
Applied Biosystems StepOne
™
and StepOnePlus
™
Real-Time PCR Systems
Installation, Networking, and Maintenance Guide
CONFIDENTIAL — For AB Internal Use Only. Do Not Distribute.
baseline-corrected
normalized reporter
(
∆Rn)
The magnitude of normalized fluorescence signal generated by the reporter:
1. In experiments that contain data from real-time PCR, the magnitude of normalized
fluorescence signal generated by the reporter at each cycle during the PCR
amplification. In the
∆Rn vs. Cycle amplification plot, ∆Rn is calculated at each
cycle as:
∆Rn (cycle) = Rn (cycle) − Rn (baseline), where Rn = normalized reporter
2. In genotyping experiments and presence/absence experiments, the difference in
normalized fluorescence signal generated by the reporter between the pre-PCR read
and the post-PCR read. In the allelic discrimination plot (genotyping experiments)
and the presence/absence plot (presence/absence experiments), ∆Rn is calculated
as:
∆Rn = Rn (post-PCR read) − Rn (pre-PCR read), where Rn = normalized reporter
See also normalized reporter (Rn).
blocked IPC In presence/absence experiments, a reaction that contains IPC blocking agent, which
blocks amplification of the internal positive control (IPC). In the StepOne
™
software, the
task for the IPC target in wells that contain IPC blocking agent. See also negative control-
blocked IPC wells.
calibrator See reference sample.
chemistry See reagents.
colocated layout A system layout in which the StepOne
™
or StepOnePlus
™
instrument is directly
connected to a colocated computer by the yellow cable. In this layout, you can control the
instrument with the StepOne
™
software on the colocated computer or with the instrument
touchscreen.
comparative C
T
(
∆∆C
T
) method
Method for determining relative target quantity in samples. With the comparative C
T
(
∆∆C
T
) method, the StepOne
™
software measures amplification of the target and of the
endogenous control in samples and in a reference sample. Measurements are normalized
using the endogenous control. The software determines the relative quantity of target in
each sample by comparing normalized target quantity in each sample to normalized target
quantity in the reference sample.
C
T
See threshold cycle (C
T
).
custom dye Dye that is not supplied by Applied Biosystems. Custom dyes may be adapted for use in
experiments on the StepOne
™
and StepOnePlus
™
systems. When using custom dyes, the
custom dye should be added to the Dye Library and a custom dye calibration performed.
IMPORTANT! Applied Biosystems does not recommend the use of TAMRA
™
dye as
reporter or quencher with the StepOne
™
system. TAMRA dye may be used as a reporter
or quencher with the StepOnePlus
™
system.
cycle threshold See threshold cycle (C
T
).