Agilent Technologies 4150 TapeStation system User Manual

202 Agilent 4150 TapeStation System Manual
10 Troubleshooting
Troubleshooting RNA Applications
Missing Marker Peaks
Incorrect protocol
used
Differences exist between Standard and High
Sensitivity assay protocols.
Ensure that the correct sample protocol for the
application is used, “Reagents and Reagent
Mixes” on page 86.
Sample prepared too
long before analysis
Evaporation or settling may alter the sample to
buffer ratio.
Ensure that your run is started immediately
after sample preparation.
The sample/buffer mix may evaporate over
time if a 96-well sample plate was used which
was not sealed with the recommended seal foil.
Always cover with the recommended foil seal
(PN 5067-5154) and ensure tight sealing
throughout the 96-well sample plate.
Incorrect reagents
used
Reagents from different assays may differ in
dye and marker concentration. Furthermore
markers used are different and upper markers
may not be present.
Ensure that the correct reagent buffer for the
application is used, see “Reagents and Reagent
Mixes” on page 86.
Table 12 RNA - Possible causes of missing marker peaks in order of probability
Cause Explanation Solution
Sample
concentration
outside
recommended range
for application
The majority of the dye binds to the sample so
that the signal from the marker is too weak to
be detected if the sample is too concentrated.
Dilute your sample until it is within the
recommended range for the application as
stated in the assay Quick Guide, see Ta ble 3 on
page 64; then prepare a new run, see
“Quantitative range” on page 107.
Insufficient migration Marker peak may not be completely resolved
from a sample peak or is not at an expected run
distance.
To check migration true distance, unalign the
gel image using the Aligned button. See
“Unexpected Migration Profile” on page 205,
“Maximum Buffer Concentration in Sample” on
page 110, and “Quantitative range” on
page 107.
Blank lane See the Agilent Information Center for more
information.
Table 11 RNA - Possible causes of incorrect quantification in order of probability.
Root cause Explanation Solution

Agilent Technologies 4150 TapeStation system Specifications

General

Brand	Agilent Technologies
Model	4150 TapeStation system
Category	Laboratory Equipment
Language	English