YouTube Transcript: How to make an acrylamide gel for electrophoresis | YouTubeToText
YouTube Transcript: How to make an acrylamide gel for electrophoresis
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when placing the glass in the casting
frame make sure it's on a flat surface
so it'll seal tightly when placed in the
casting stand
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you may want to Mark the glass as a
guide so you know how much running gel
to add ideally Your Mark should be about
1 cm below the teeth of the comb you can
insert the comb to double
check once the APS and tet are added the
solution will start to polymerize so try
to add them as quickly as possible
[Music]
to add the running gel solution place
the tip of the PIP pet in the corner of
the space between the short plate and
the spacer plate it's a good idea to
leave a little bit of solution in the
pipet to avoid introducing bubbles into
the gel continue filling until the
solution reaches Your
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Mark adding isopropanol to the top of
the gel will keep the solution from
forming a meniscus within the glass
resulting in a very smooth surface don't
forget to remove the isopropanol layer
before adding the stacking gel
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once the running gel solution has
polymerized and formed a plug you may
add the APS and Ted to the stacking gel
solution and then add it to the top of
the cassette typically you should allow
20 minutes for polymerizing
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ation be careful not to damage the gel
while removing the isopropanol layer
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remember that APS and Ted cause the
solution to polymerize so add them
immediately proceeding addition to the
cassette
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bring the level of the stacking gel
solution to the top of the short glass
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when inserting the comb some of the
stacking gel solution will spill over
wait until the solution is polymerized
before cleaning the acrylamide is
considered toxic until polymerization
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occurs when a gelatinous plug forms the
vile you are ready to remove the comb
from the
gel the wells can be a little tricky to
find Once the comb has been removed from
the finished gel so marking the first
well prior to removing the comb can save
you some
time be careful not to remove the comb
at an angle because this can damage the
wells
[Music]
you are now ready to run your gel
through electropheresis to examine your
sample good luck
[Music]
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