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Caution: Ethidium bromide
is a known mutagen. Always
wear gloves when handling.
Wear UV safety goggles and
protect skin when using a UV
lamp.
Note: If no dye was added to
the coolant, place the base on
a dark background to see the
wells more easily.
Note: At the maximum setting
the unit begins overheating
as soon as the chilled base
reaches ambient temperature.
If overheating is not
controlled, the gel will melt
and/or the base of the unit
will warp!
The electrophoresis run
Refer to the notes, buffers, and volumes section for
additional information and guidelines.
Chill the base before use, especially when higher
voltage settings will be used or when the separation
will require more than 30 minutes.
Note: To monitor separation progress, either add 0.5
µg/ml (final conc.) of ethidium bromide to the running
buffer now, or add 50 µg/ml (final conc.) ethidium
bromide to the sample buffer. To visualize progress,
turn off the power supply, remove the lid assembly,
and hold a portable UV lamp near the gel.
Adding ethidium bromide to the running or sample
buffer slows migration slightly. Detection by this
method is not as sensitive as staining and viewing on
a transilluminator. (See DNA detection, p. 15.)
Fill both buffer chambers with running buffer until
the buffer is ~1 mm deep over the gel. (This requires
about 220 ml.)
Load the samples.
Add sample to 5X sample loading buffer and mix (1/5
of the final volume is loading buffer, see p. 14). Use
a micro-pipette to load each sample, taking care to
avoid puncturing the well bottom or entrapping any
bubbles.
Place the lid so that the cathode (–, black lead) is
at the end nearest the sample well. (Nucleic acid
samples migrate toward the anode, +, red lead.)
Connect the color-coded leads (red to red, and
black to black) to an approved power supply, such
as the EPS 2A200. Set the voltage level and timer
(if available) according to the degree of resolution
sought.
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p9
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