C.B.S. Scientific MGV-402 - Manual (Page 14)

14
MGV Mini-Vertical Instructions 9-30-13
SECTION 4
Running Conditions
4.1 Recommended Power for Slab Gels:
Precise electrophoresis conditions will vary according to the number and type of
gels used, buffer conditions employed, power input, and the general goal of the
experiment. Refer to the reference section for in depth discussions on practical; and
theoretical approaches to protein gel electrophoresis.
Using standard SDS-PAGE buffer systems for a single 0.75mm thick gel at room
temperature (cat. # MGV-102 or MGV-302) use the following conditions at constant
voltage:
70VDC, 15mA, 4 hours
120VDC, 20mA, 2 hours
150VDC, 25mA, 1.5 hours
180VDC, 20mA, 1.0 hour
If running two gels in the Dual Units (cat. #MGV-202 or MGV-402), keep the volts
the same but double the mA. It is also true that if the thickness of gel increases,
increase the mA proportionally.
At constant voltage, the proteins will migrate at a constant rate during electropho-
resis with adequate heating appropriate for denaturing gels. Increasing the voltage/
mA (for a single gel thickness and percentage) will speed mobility but increase the
risk of overheating.
For systems which employ active temperature control (cat. # MGV-202 or MGV-
402) the power input, thus the migrations rate, can be increased. The joule heating
generated by the higher power is offset by the cooling effect of the water between
the gels. Exact conditions should be determined empirically but should be at least
in the 20% range.
4.1.1 ClearPAGE Gels
Run Voltage Starting Current Ending Current Approx. Run TIme
180VDC 90mA/gel 40mA/gel 30-75 minutes
4 1 2 Tris-Glycine Gels
For SDS-PAGE Tris-Glycine (Laemmli) buffer systems with two 1.0mm thick gels at
room temperature use the following conditions at constant voltage:
80VDC until samples have fully entered stacking gel
120VDC @ 60mA-90mA/gel (depending on gel type) thereafter until dye is
near bottom of gel