User guide
65
GlobalFiler
™
Express PCR Amplification Kit User Guide
A
Troubleshooting
Follow the actions recommended in this appendix to troubleshoot problems that occur
during analysis.
Table3 Troubleshooting
Observation Possible causes Recommended actions
Faint or no signal from
both the DNA Control 007
and the DNA test
samples at all loci
Incorrect volume or absence of Master
Mix or Primer Set
Repeat amplification.
No activation of DNA Polymerase Repeat amplification, making sure to hold reactions
initially at 95°C for 1 minute.
Master Mix not vortexed thoroughly
before aliquoting
Vortex the Master Mix thoroughly.
Primer Set exposed to too much light Store the Primer Set protected from light.
Evaporation. Ensure that the plate is properly sealed with film
and that you used a compression pad with the 9700
thermal cycler (a compression pad is not needed
with the Veriti
®
thermal cycler).
PCR System malfunction Refer to the thermal cycler user’s manual and
check instrument calibration.
Use of incorrect thermal cycling
parameters
Check the protocol for correct thermal cycling
parameters.
MicroAmp
®
Base used with tray/
retainer set and tubes in GeneAmp
®
9700
Remove MicroAmp Base from tray/retainer set and
repeat test.
Insufficient PCR product
electrokinetically injected
Prepare PCR product as described in Chapter3,
“Perform Electrophoresis” on page 29.
Degraded formamide Check the storage of formamide; do not thaw and
refreeze multiple times. Try Hi-Di™ Formamide.
Sample punch location was not
optimal
For blood samples on treated paper, punch in the
center of the blood stain.
For buccal samples on treated paper, punch in the
center of the buccal transfer or punch in the
optimal spot based on past experiences.
For buccal samples collected with the Bode Buccal
DNA Collector
™
, punch from near the tip of the
collector.
Insufficient lysis of the swab head Ensure swab heads are incubated for 20 minutes in
400 µL Prep-N-Go
™
buffer.