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0:05
Finally, I would
just like to show
some selected applications
of capillary gel
electrophoresis.
0:13
This slide shows the very
first capillary gel
electrophoresis of
DNA fragments of sizes
between the 10 mers, 30
mers, the 40 and 60 mers.
This was the basis of DNA
sequencing in the old days
in capillary electrophoresis
because it showed that there is
an option to use
capillary columns to
separate of DNA fragments
differing only by
one nucleotide.
In this case,
picomolar amounts
of a mixture of
these polydeoxyadenylic acids,
the baseline was solved in
less than 11 minutes by
capillary gel electrophoresis
in polyacrylamide gels.
0:55
Even more, the
separation pair was
demonstrated in this
figure than those are
non-denaturing capillary
polyacrylamide gel
electrophoresis separations
of three human K-ras oncogene
probes differing only by
one nucleotide in the
middle of the molecule as
one can see within
less than 30 minutes;
not baseline because
of the length of
the capillary but all
of these three were
separated again, only having
one difference in the middle
of the exact same sequence.
1:32
That was the basis
of DNA sequencing in
the early days when
the second- and third-generation
sequencers were not
available yet so very long
read length capillary
gel electrophoresis
base sequencing was
possible to use
capillary chiller.
Choosing the appropriate
parameter separation
voltage and temperature,
high-resolution DNA
sequencing prep played
an important role in obtaining rapid
and high resolution separation.
The sequencing speed of more
than 1,000 bases per
run was successfully
achieved by this one method and
increased separation temperature
which has multiple
benefits such as
speeding up the separation
and increasing the accuracy and
the read length
and additionally,
providing better denaturation.
