Bio-Rad Experion RNA Analysis Kits - Manual (Page 49)

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Error Probable Cause Recommended Action
Lower marker is identified
incorrectly Experion software selected incorrect peak to
correspond to the marker; such an error affects
alignment and sizing Manually select the markers by following the instructions in
Section 6.1
Virtual gel does not
resemble an agarose gel
separation of same samples Gel view is a direct interpretation of
electropherogram data. By default,
fluorescence intensity of bands in each lane is
scaled to intensity/peak height of highest peak
in that sample (scale to local). Consequently,
each gel lane may have a different scale of
fluorescence intensity Adjust scaling of the virtual gel to Scale To Global (see
Section 4.2). This adjusts the scale on all electropherograms
Adjust the intensity of the virtual gel using the sliding cursor
Baseline is fluctuating (for
example, into very negative
and positive values) Air bubbles are interfering with electrical
contact in one or more of the wells Stop the run, remove the chip, and remove and replace
solution in affected wells
Sample is in inappropriate sample buffer Ensure RNA is in buffer of low ionic strength (for Experion
RNA StdSens use TE or water, for Experion RNA HighSens
use water)
Chip is not completely primed Prime and load a new chip
Samples are too concentrated; RNA levels are
out of range of assay Dilute the samples and repeat the analysis
RNA ladder or samples
appear degraded RNase contamination of electrodes, reagents,
or consumables
Improper storage
Perform the deep cleaning procedure described in
Appendix B and software Help section (search term
“electrodes”)
Use only 0.2 µm-filtered water (such as Experion DEPC-
treated water) for diluting samples and the RNA ladder
Use RNase-free consumables and RNase-free, filter pipet
tips; change pipet tips with each use
Store RNA ladder and samples appropriately (for example,
at –70°C)
7.3 Contacting Technical Support
If you are having problems with a run and would like the Bio-Rad Technical Support team to review your
data, please submit the following files to LSG_TechServ_US@bio-rad .com for analysis:
n
Run file (C:\Program Files\Bio-Rad Laboratories\Experion Software\Data\
name>\.bdf)
n
Run log (C:\Program Files\Bio-Rad Laboratories\Experion Software\Data\
name>\.log)
n
Packet file (C:\Program Files\Bio-Rad Laboratories\Experion Software\Data\
name>\Packet\.txt)
n
System log (C:\Program Files\Bio-Rad Laboratories\Experion Software\Backup\Version X\
Experion_System.log or .txt)
n
Lot numbers of the RNA chips and the Experion RNA analysis kit
To export the run file, log file, and packet file, select the run(s) in the tree view and click Export Selected
Runs. The software exports the files into folders with the corresponding project name(s).
Experion RNA StdSens and HighSens Analysis Kits