Pure Dye Runs - Applied Biosystems 7900HT User Manual

Pure Dye Runs

Pure Dye
Troubleshooting
Table
Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
6. Repeat step 4 until you identify the location of each contaminated well.
7. Decontaminate the sample block as explained in
Block" on page 7-14.
8. Run a background plate to confirm that the contaminants have been removed.
If the contamination is present after running the background plate for a second
time, the background plate is likely to be the source of contamination.
Table 8-3 Troubleshooting Pure Dye Runs
Observation
Software will not extract
pure dye data
Raw data from pure dye
run appears strange
(see below)
Signals plateau (saturation)
Signal is too low
(< 10,000 FSU)
More than two outliers per
dye in a single row
Possible Cause
During plate setup, the
wrong plate type was
assigned to the plate
document
A background plate was not
run before the pure dye
plate or card
Pure dye plate or card was
loaded backwards
Intensity is set too high/low
• Evaporation
• Contamination
DRAFT
September 1, 2004 11:39 am, CH_Trouble.fm

Pure Dye Runs

"Decontaminating the Sample
Recommended Action
Create and run a new pure
dye plate document with the
proper plate type setting
Run a background plate,
then run the pure dye plate
or card again
1. Verify the pure dye
wavelengths are as
expected.
2. Rerun the pure dye plate.
or card
Call Applied Biosystems
Technical Support.
Rerun the pure dye plate or
card. If the problem
persists, discard the pure
dye plate or card and run a
new one.
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