Hoefer SE615 - Manual (Page 14)

Pouring standard gradient gels
See Fig 1 on page 4 for a diagram of the caster
components. See “Setting up the caster” on
page 5 for recommended solution volumes.
Setting up the gradient maker
1
Prepare a 50% glycerol displacement solution
containing a small amount of bromophenol blue:
approximately 75 ml for the SE615 and 35 ml for the
SE675.
2
Calculate the amount of gradient solution you will
need by referring to the formula on page 9.
For one-dimensional gels: Use a 12-cm separation
gel with a 4-cm stacking gel.
For two-dimensional gels using IPG strips: Calculate
the amount of solution needed to fill the sandwiches
to approximately 0.5–1.0 cm below the top of
the plates.
3
Set up the peristaltic pump. Using a graduated
cylinder and water, adjust the flow rate so that the
volume of the gradient separation solution plus the
volume of the glycerol solution will be completely
delivered in 10 –15 minutes.
4
Choose a gradient maker that holds no more than
four times the total volume of gradient solution to
be poured. Check that the gradient maker is clean
and that the outlet and mixing port are free of
polymerized acrylamide.
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