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R
C
EMOVING THE
OMB
1.) When the gel is solidified and fully opaque, carefully remove the comb with a gentle wiggling,
upward motion. If the comb is difficult to remove or if a low percentage gel is being used,
overlay the comb area with a small volume of 1X electrophoresis buffer to preserve the integri-
ty of the wells. Check the wells to ensure their bases are intact.
CAUTION: Prolonged exposure of the Delrin combs to gels containing formaldehyde will cause
them to degrade. Be sure to remove the comb(s) from formaldehyde gels as soon as
gel hardening is complete and rinse them well prior to storage.
If a gel is not to be used immediately after preparation, remove it from the casting fixture and place
it in a plastic bag or container submerged in 1X electrophoresis buffer containing 1mMNaN3. Store
at +4 o C.
L
S
OADING THE
AMPLES INTO THE
1.) Remove the casting tray containing the hard-
ened agarose gel from the casting fixture by
pressing the casting tray against the foam pad
and lifting at the thin pad end. Place the tray
and gel into the main unit assembly such that
the samples wells are on the same end as the
negative (black) electrode. (see Photo 3)
2.) Fill the unit with the remaining 1X elec-
trophoresis buffer containing ethidium bro-
mide made previously (or 1X MAE buffer
for RNA gels), covering the gel to a depth of
1-5mm. Approximately 250ml of buffer will
be required.
NOTE: Use of the same batch of electrophoresis
buffer for both the gel and the running
buffer is very important. Slight variations in buffer composition between gel and running
buffer may result in ionic or pH gradients that can significantly impact the mobility of
the samples.
3.) Pre-run RNA gels at 100V for five minutes prior to loading the samples.
4.) Load the samples into the wells with a micropipette or similar device taking care not to punc-
ture the bottom of the wells or load the sample onto the top of the gel. For improved well visu-
alization during sample loading, be sure that the wells are positioned over the contrasting
stripes located on the bottom of the buffer tanks.
G
EL
Photo 3
7
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