Applied Biosystems StepOne User Manual

DRAFT September 25, 2007 1:07 am, 4376782_Troubleshoot.fm
Appendix A How to Troubleshoot the Installation
Dye Calibration Failure
135
Applied Biosystems StepOne
™
and StepOnePlus
™
Real-Time PCR Systems
Installation, Networking, and Maintenance Guide
Notes
Common
Irregularities
The following table displays irregular spectra common to dye calibrations.
Spectra Possible Cause Action
One or more raw spectra are
at or below the detectable
threshold for the calibration.
• The spectral calibration plate has
been centrifuged insufficiently.
•The spectral calibration plate
contains old or insufficient reagents.
• If you are running a custom spectral
calibration plate
, the dye may not
be present at a sufficient
concentration.
1. Unload the instrument and view the wells of the
spectral calibration plate . If the liquid in the wells
are not:
– At the bottom of the wells, centrifuge the plate
for a longer time, then repeat the calibration.
– Equivalent in volume, the plate is not sealed
and the reagents have evaporated. Discard it
and run another.
2. If the spectral calibration plate appears to be
normal, discard the plate and run another.
3. If the problem persists, contact
Applied Biosystems as explained in “How to
Obtain Support” on page x.
Note: If you are running a custom spectral
calibration plate, create another plate but increase
the concentration of the dye that produced
insufficient signal.
One or more raw spectra
exceed the maximum limit for
the instrument.
• Fluorescent contamination is
present on the sample block(s) or
spectral calibration plate.
• If you are running a custom spectral
calibration plate
, the dye may be
too concentrated.
Confirm that the presence of contamination by
performing a background calibration as explained
in “Perform a Background Calibration” on page 94.
If the background calibration does not show
sample block contamination, the spectral
calibration plate may be contaminated.
Note: If you are running a custom spectral
calibration plate, create another plate but decrease
the concentration of the dye that exceeds the
detectable limit.
The spectra contain peaks in
more than one filter.
Fluorescent contamination is present
on the sample block(s) or spectral
calibration plate.

Applied Biosystems StepOne Specifications

General

Brand	Applied Biosystems
Model	StepOne
Category	Laboratory Equipment
Language	English