Hoefer HE-PLUS Manual Del Usuario página 16

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.
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p6
C. Removing the Comb
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 integrity of the wells. Check the wells to
ensure their bases are intact.
D. Loading the Samples onto the Gel
1
Remove the casting tray containing the hardened
agarose gel from the casting fixture by lifting the
ends. Place the tray and gel into the main unit
assembly such that the sample wells are on the same
end as the negative (black) electrode.
2
Fill the unit with the remaining 1X electrophoresis
buffer containing Ethidium Bromide made previously,
covering the gel to a depth of 1– 5 mm. Approximately
350 ml of buffer will be required.
3
Load the samples into the wells with a micropipette or
similar device taking care not to puncture the bottom
of the wells or load the sample onto the top of the gel.