200 Agilent 4150 TapeStation System Manual
10
Troubleshooting
Troubleshooting RNA Applications
Troubleshooting RNA Applications
Incorrect Quantification Results
Quantification calculation is facilitated by comparing the area under a peak or
region of interest in the electropherogram with the area under the Lower
Marker. The Lower Marker has a defined concentration.
Incorrect quantification results can have multiple causes. If, after checking
each of these causes, your results files continue to show concentration
discrepancies, please contact your local support representative.
The troubleshooting information is complemented by How To videos or animated graphics
which are provided in the Agilent Information Center (see “Agilent Information Center” on
page 60).
Table 11 RNA - Possible causes of incorrect quantification in order of probability.
Root cause Explanation Solution
Insufficient Mixing Buffer and sample was not sufficiently
vortexed. The dye could not incorporate
completely into the sample.
See “Mixing Recommendations” on page 94
and the Agilent Information Center for more
information.
Sample
concentration
outside
recommended range
for application
Linear concentration estimation cannot be
achieved for samples exceeding the limits in
the specification table.
Either dilute or concentrate the sample until it
is within the recommended range for the
application as stated in the respective assay
Quick Guide; then prepare a new run. Use the
High Sensitivity assay if applicable
(“Quantitative range” on page 107, Tab le 3 on
page 64).
Incorrect pipetting
technique or pipette
calibration
Incorrect pipetting technique or uncalibrated
pipettes can lead to poor pipetting performance
which in turn alter sample/buffer ratio in an
unpredictable manner.
See “Good Pipetting Practice” on page 85 and
follow guidelines of the respective pipette
vendor.
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