Bio-Rad Experion DNA Analysis Kits - Manual (Page 44)

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7.1 Electrophoresis and Priming Stations
Error Probable Cause Recommended Action
IV Check Failure
(Experion
™ software)
One or more wells are not filled correctly, and
the electrodes are not immersed Fill all sample wells of the chip with blank or replicate samples
Ensure pipets are properly calibrated
Air bubbles are interfering with the electrical
contact in one or more of the wells Stop the run, remove the chip, and use a clean pipet tip to
remove the sample and apply a new aliquot
When pipetting, insert the tip vertically and to the bottom of
the well. Dispense the liquid slowly. Do not expel air at the end
of the pipetting step. Dispense only to the first stop on the
pipet. Positive (reverse) pipetting may also be used
An electrode is loose or disconnected Perform a conductivity test with the cleaning chip or
conductivity chip from the Experion validation kit (catalog
#700-7051). Add 850 µl water to the chip, place the chip in
the electrophoresis station, and close the lid. In Experion
software, select Tools > Diagnose Instrument and select
Conductivity in the Instrument Diagnostics dialog. If the
test passes, focus on the sources of error above. If it fails,
contact Bio-Rad Technical Support with the results
Lid is not closed Close the lid
Chip is not primed properly Repeat the analysis using a new chip
Check Seal
(Priming station) Priming seal is worn, torn, or clogged Replace the priming seal (catalog #700-7031)
Chip is not positioned correctly in the priming
station Align the arrow on the chip with the arrow on the platform
Gel-stain solution (GS) was added to the
wrong well for priming The priming well is highlighted on the chip. See Section 3.5,
Prime the Chip
No chip detected Chip not positioned properly, empty wells,
or incorrect reagents Make sure the chip is positioned properly and correct
reagents are used
Run aborts before all wells
are analyzed Computer power saver options are causing
runs to abort (this is a problem only with
Experion software prior to version 3.0) Upgrade to the newest software version
Confirm that all power saver and/or power options are turned
off before starting the assay. Do not let the computer “sleep”
during a run. If the computer goes to sleep, the run aborts
7.2 Experion DNA Analysis
Error Probable Cause Recommended Action
Concentrations are not
reproducible (CV > 20%) Sample preparation procedures are
inconsistent Use the same reagents and follow the same procedures
for each sample and each run. Do not modify the protocol
outlined in Chapter 3 to compensate for DNA concentration
or sample variability
Peaks were not detected in
the DNA ladder An air bubble is at the bottom of the ladder
well or microchannels due to poor priming or
sample loading; air bubbles can interfere with
the contact between the electrode and the
liquid Check the chip for air bubbles after priming and sample
loading:
n After priming: turn the chip over to make sure the
microchannels are filled with gel-stain solution (GS).
If the chip is primed correctly, the wells should be hard to
see. Compare the primed chip to an unprimed chip
if necessary. Do not touch the glass
n After sample loading: tilt the chip and view it from
different angles against a light background
Experion Automated Electrophoresis System