Applied Biosystems StepOne User Manual

DRAFT September 25, 2007 1:07 am, 4376782_Maintain_the_System.fm
Chapter 6 Maintain the System
Regular Maintenance
Applied Biosystems StepOne
™
and StepOnePlus
™
Real-Time PCR Systems
Installation, Networking, and Maintenance Guide
100
Notes
Perform a Dye Calibration
Perform a dye calibration at least every 18 months, or as often as necessary depending on
instrument use. Because the age and use of the instrument can affect spectral readings,
you may need to perform the calibration more frequently.
IMPORTANT! Perform a background calibration before performing a dye calibration (see
“Perform a Background Calibration” on page 94).
Materials
Required
• Spectral Calibration Plates from the spectral calibration kit for your instrument:
– StepOne
™
Real-Time PCR System Spectral Calibration Kit (PN 4371433)
– StepOnePlus
™
Real-Time PCR System Spectral Calibration Kit (PN 4371435)
• Safety glasses
• Powder-free gloves
• Centrifuge with reaction plate adapter
• StepOne system cable, yellow (from the system packing kit)
Custom Dye Plates
The StepOne
™
and StepOnePlus
™
instrument supports the use of custom dyes (dyes not
supplied by Applied Biosystems) that fluoresce within the spectral range it supports (500
to 650 nm). See “How to Calibrate a Custom Dye” on page 110 for more information.
Purpose of the
Calibration
During a dye calibration run, the system:
• Collects spectral data from a series of dye standards.
• Stores the spectral information for the dye standards in the spectra run file, a
calibration file in the StepOne
™
software directory.
The StepOne
™
software uses spectral data during subsequent runs to characterize dyes
and distinguish the individual contribution of each dye in the collective fluorescence
collected by the instrument during a run. After each run, the StepOne
™
software stores
run data in the form of a raw spectral signal for each reading. It determines the
contribution of each fluorescent dye used in the sample by comparing the raw spectra to
the spectral file. After analysis, the StepOne
™
software stores the spectral data with the
collected fluorescence data for each experiment.

Applied Biosystems StepOne Specifications

General

Brand	Applied Biosystems
Model	StepOne
Category	Laboratory Equipment
Language	English