Thermo Scientific appliedbiosystems SeqStudio Flex Series - Page 515

Observation Possible cause Recommended action
No signal or a low signal is
detected
Note: Samples are stable
on the instrument for 16–24
hours. Determine if samples
will be stable if a re-injection
is recommended, then plan the
re-injection accordingly.
(continued)
The Hi‑Di
™
Formamide used
to prepare the samples was
degraded.
Prepare the samples with fresh Hi‑Di
™
Formamide and repeat the experiment.
See “Sample preparation guidelines” on
page 73.
The sample was prepared
with the BigDye XTerminator
™
Purification Kit but a BDX run
module was not selected.
Re-inject the sample with the correct module.
The sample was degraded. Prepare the sample according to the protocol
provided with the kits for sample preparation.
See “Sample preparation guidelines” on
page 73.
IMPORTANT! Do not r
esuspend samples in
water.
Thermal breakdown of samples is normal.
Samples are stable for 16–24 hours on the
instrument.
Use Hi‑Di
™
Formamide to prepare the
samples. Sample stability is optimal in Hi‑Di
™
Formamide.
The sample had a high
concentration of salt, or
There was an excess of
unlabeled template competing
with the fragments labeled with
dye during the injection.
Dilute or desalt the samples.
For more information, see the following
publications:
•
DNA Sequencing by Capillary
Electrophoresis Chemistry Guide Second
Edition (Pub. No. 4305080)
•
DNA Fragment Analysis by Capillary
Electrophoresis User Guide (Pub.
No. 4474504)
A capillary array tip is blocked. Run the Fill array with polymer maintenance
wizard. (In the home screen, tap
Actions4Maint
enance4Maintenance4
wizards4Fill array with polymer.)
Replace the capillary array if the problem
persists.
The capillary array is damaged. Inspect the capillary array for damage.
Replace the capillary array if there is damage.
If replacing the capillary array does not resolve
the issue, and the issue is occurring in all
capillaries, contact Technical Support.
Pull-up or pull-down peaks are
pr
esent in the data
The incorrect dye set was
selected in the plate file.
Correct the dye set and repeat the injection.
Appendix A Troubleshooting
Sample and data troubleshooting
A
SeqStudio
™
Flex Series Genetic Analyzer with Instrument Software v1.0 User Guide
515