Agilent 4150 TapeStation System Manual 203
Troubleshooting
10
Troubleshooting RNA Applications
Degraded RNA Ladder and/or Samples
Table 13 RNA - Possible causes of degraded sample in order of probability
Root cause Explanation Solution
Incorrect heating
step
If heat denaturation was too long and/or too
high temperatures were used the sample will
degrade upon this treatment
Ensure the correct Heat Denaturations for the
assay are followed as described in the
appropriate Quick Guide, then prepare a new
run(“Heat Denaturation” on page 99, Ta ble 3 on
page 64)
Sample
concentration
outside
recommended range
for application
A too concentrated sample will significantly
alter the electropherogram
Either dilute or concentrate your sample until it
is within the recommended range for
application as stated in the assay Quick Guide,
then prepare a new run(“Quantitative
range” on page 107)
Insufficient migration
due to presence of
salt
Sample matrices exceeding the maximum
buffer strength specifications may alter
currents in an unpredictable manner leading to
short running lanes. See “Maximum Buffer
Concentration in Sample” on page 110.
Dilute your samples to ensure low levels of
buffer salt. True migration profiles can be seen
by unaligning your gel image by pressing the
Aligned button in the TapeStation Analysis
software. Please refer to the salt tolerance
guidelines for your assay in the appropriate
Quick Guide (Tab le 3 on page 64)
RNAse
contamination of the
reagents
RNAse contamination will lead to degradation
of the sample
Ensure Good Measurement Practices when
working with RNA assays; then prepare a new
run using new reagents (“Good Measurement
Practices” on page 81).
RNAse
contamination of the
plasticware
RNAse contamination will lead to degradation
of the sample.
Ensure Good Measurement Practices when
working with RNA assays; then prepare a new
run using new reagents (“Good Measurement
Practices” on page 81).
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