Applied Biosystems StepOnePlus User Manual

DRAFT September 25, 2007 1:07 am, 4376782_Glossary.fm
Glossary
Applied Biosystems StepOne
™
and StepOnePlus
™
Real-Time PCR Systems
Installation, Networking, and Maintenance Guide
CONFIDENTIAL — For AB Internal Use Only. Do Not Distribute.
144
primer/probe mix PCR reaction component that contains the primers designed to amplify the target and a
TaqMan
®
probe designed to detect amplification of the target.
pure dye See custom dye and system dye.
quantitation
method
In quantitation experiments, the method used to determine the quantity of target in the
samples. In StepOne
™
and StepOnePlus
™
systems, there are three types of quantitation
methods: standard curve, relative standard curve, and comparative C
T
(∆∆C
T
).
quantity In quantitation 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.
quencher A molecule attached to the 3′ end of TaqMan
®
probes to prevent the reporter from
emitting fluorescence signal while the probe is intact. With TaqMan
®
reagents, a
nonfluorescent quencher-minor groove binder (NFQ-MGB) can be used as the quencher.
With SYBR
®
Green reagents, no quencher is used.
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.
QuickStart A feature in StepOne
™
and StepOnePlus
™
systems that allows you to run an experiment
without entering plate setup information. QuickStart requires a colocated layout with the
instrument powered on and an intact instrument-computer connection.
R
2
value Regression coefficient calculated from the regression line in the standard curve. 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.
ramp The rate at which the temperature changes during the instrument run. Except for the melt
curve step, the ramp is defined as a percentage. For the melt curve step, the ramp is
defined as a temperature increment. In the graphical view of the thermal profile, the ramp
is indicated by a diagonal line.
ramp speed Speed at which the temperature ramp occurs during the instrument run. Available ramp
speeds include fast and standard.
• For optimal results using the fast ramp speed, Applied Biosystems recommends
using TaqMan
®
Fast reagents in your PCR reactions.
• For optimal results using the standard ramp speed, Applied Biosystems
recommends using standard reagents in your PCR reactions.
IMPORTANT! TaqMan Fast reagents are not supported for genotyping or
presence/absence experiments.
raw data plot A plot of raw fluorescence signal (not normalized) for each optical filter.

Applied Biosystems StepOnePlus Specifications

General

Brand	Applied Biosystems
Model	StepOnePlus
Category	Laboratory Equipment
Language	English