DRAFT
September 25, 2007 1:07 am, 4376782_Maintain_the_System.fm
Chapter 6 Maintain the System
Regular Maintenance
101
Applied Biosystems StepOne
™
and StepOnePlus
™
Real-Time PCR Systems
Installation, Networking, and Maintenance Guide
Notes
Supported Dyes
StepOnePlus
™
System Dyes
The StepOnePlus
™
instrument uses the following dye set for calibration: FAM
™
dye,
JOE
™
dye, NED
®
dye, ROX
™
dye, TAMRA
®
dye, VIC
®
dye, and SYBR
®
Green dye.
The following figure shows the emission spectrum for each dye, and the filter at which
each dye is read.
StepOne
™
System Dyes
The StepOne
™
instrument uses the following dye set for calibration: FAM
™
dye,
JOE
™
dye, ROX
™
dye, VIC
®
dye, and SYBR
®
Green dye. The following figure shows
the emission spectrum for each dye, and the filter at which each dye is read.
Custom Dyes
The system can be used to run assays designed with custom dyes (dyes not supplied by
Applied Biosystems). However, before using custom dyes with the instrument, you must
create and run a custom calibration plate. The purpose of the custom plate is similar to
that of a spectral calibration plate. The StepOne
™
software uses the custom
calibration plate to create a spectral standard to distinguish the custom dye in the
fluorescence data collected during the run.
IMPORTANT! To use a custom dye on your system, it must fluoresce within the 500 to
650 nm spectral range measured by the StepOne
™
and StepOnePlus
™
instrument.
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.
31Filters 4
Wavelength
(nm)
Emission
Spectra
Dyes
2
~520 nm
• FAM
• SYBR Green
• JOE
• VIC
• ROX
~610 nm
• NED
• TAMRA
~580 nm~550 nm
500 600 700
Filters 1 2 3
Wavelength
(nm)
Emission
Spectra
Dyes
~520 nm
• FAM
• SYBR Green
• JOE
• VIC
• ROX
~610 nm~550 nm
500 600 700
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