Thermo Scientific appliedbiosystems SeqStudio Flex Series - Sequencing Applications

Observation Possible cause Recommended action
Poor resolution in all the
capillaries
(continued)
The Hi‑Di
™
Formamide that
was used to prepare the
samples was degraded.
Prepare the samples with fresh Hi‑Di
™
Formamide and repeat the reaction.
See:
•
“Sample preparation guidelines” on
page 73
•
DNA Fragment Analysis by
Capillary Electrophoresis User Guide
(Pub. No. 4474504)
Sample contains high amount
of DNA.
Dilute the sample and re-inject it.
Problem with capillary array. Run the Change capillary array maintenance
wizard to uninstall, then re-install the
capillary array. (In the home screen, tap
Actions4Maintenance4Maintenance
wizards4Change capillary array.)
Sequencing applications
Observation Possible cause Recommended action
Short read length and uneven
peak spacing is observed in
the sequence data
The incorrect dye set was
selected in the plate file.
Correct the dye set and repeat the injection.
Select from the following:
•
E_BigDye
™
Terminator v1.1
•
Z_BigDye
™
Terminator v3.1
•
Z_BigDye
™
Direct
Reanalyze the data in the Sequencing Analysis
Software, using the correct mobility file.
Dye blobs are seen in the
sequencing data
Impurities remained in the
sample after the sample
purification. The impurities
caused dye blobs to appear in
the sequencing data.
Improve the sample purification method. See
“Sample preparation guidelines” on page 73.
Extra peaks are present in the
sequencing traces
There was renaturation of the
sample.
Heat-denature the samples prepared with
fresh Hi‑Di
™
Formamide, then immediately
place the samples on ice.
Appendix A Troubleshooting
Sample and data troubleshooting
A
SeqStudio
™
Flex Series Genetic Analyzer with Instrument Software v1.0 User Guide
517