Thermo Scientific appliedbiosystems SeqStudio Flex Series - Page 74

(continued)
Item Guidelines
Sequence analysis sample
preparation
•
Prepare sequencing reactions according to kit instructions, and purify the
extension products with ethanol precipitation, spin columns, or the BigDye
XTerminator
™
Purification Kit.
–
If ethanol precipitation or spin columns are used, dry the samples in a
vacuum centrifuge without heat or at low heat for 10–15 minutes or until
dry.
Note: Do not over dry the samples.
–
Resuspend in 10–20 µL of Hi‑Di
™
Formamide.
•
Use a 65 µL or 130 µL sample volume for samples that are prepared with the
BigDye XTerminator
™
Purification Kit. See BigDye XTerminator
™
Purification Kit
User Guide (Pub. No. 4374408).
•
Use a 10–20 µL sample volume for samples that are prepared with Hi‑Di
™
Formamide.
•
Samples prepared with Hi‑Di
™
Formamide are stable for 24 hours on the
instrument.
Note: Data quality may be reduced if samples exceed 24 hours on the
instrument. The software displays an alert if a sample will exceed 24 hours on
the instrument by the end of the run.
•
Ensure that Hi‑Di
™
Formamide is fresh.
–
Hi‑Di
™
Formamide should not undergo more than two freeze-thaw cycles
(one to aliquot and one for use).
–
Use the same day after thawing.
•
Do not resuspend samples in water, which can decrease sample stability and
create signal variability upon electo-kinetic injection.
•
For more information, see DNA Sequencing by Capillary Electrophoresis
Chemistry Guide Second Edition (Pub. No. 4305080).
Chapter 3 Prepare the samples
Sample preparation guidelines
3
74
SeqStudio
™
Flex Series Genetic Analyzer with Instrument Software v1.0 User Guide