Hoefer HE99X - Manual (Page 11)

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p3
Operating instructions
Agarose gels are first cast in the gel casting kit,
and samples are then loaded into the wells and
electrophoretically separated. The fluorescent
dye ethidium bromide can be added to the gel
or electrophoresis buffer or both in order to
track separation progress. At the completion
of electrophoresis, the gel may be stained and
photographed, blot transferred, or dried for
autoradiography.
Casting the gel
Prepare the solutions
1
Prepare about 1.3 liters of running buffer. Up to 100 ml
of buffer is required for the gel and 1.2 liters for the
buffer chamber. Refer to page 10 for recipes of three
commonly used electrophoretic running buffers.
2
Prepare the sample loading buffer. Refer to page 12
for a recipe and tabulated volume capacity for each
comb size.
3
Prepare agarose solution(s).
Dissolve agarose in running buffer, heat according to
instructions accompanying the agarose, and allow the
solution to cool to 50 °C before pouring into the
running tray.
Optional: Add 0.5 µg/ml ethidium bromide to the gel
solution in order to facilitate observation of separation
progress during electrophoresis.
Before you start…
1. Wash all components
with a dilute solution of
laboratory detergent and rinse
thoroughly.
2. Level the unit by placing the
spirit level on the running
platform and adjusting the
leveling feet.
Volume for 3-mm thick gels
tray size (cm) agarose (ml)
15 × 10 45
15 × 15 68
15 × 20 90
Caution! Ethidium bromide is
a known mutagen. Always wear
gloves when handling.