Hoefer SE215 - Manual (Page 12)

gel caster required volume gradient maker
range (ml) size*
SE215 55–100 SG100
SE235 32–56 SG50 or SG100
SE275 22–40 SG30 or SG50
*Actual size depends on calculated volume.
4
Prepare solutions
Plug solution
Prepare enough plug solution (50% sucrose or 75%
glycerol in 1X separating gel buffer) to fill the space
below the sandwiches. The SE215 requires about
30 ml and the SE235 or SE275 requires about
15 ml. The plug solution is pumped in after the gradi-
ent to displace all of the monomer solution into the
sandwich stack.
Optional: Add bromophenol blue to help track the
plug as it is pumped into the multiple gel caster.
Monomer solution
Follow the instructions on page 6, step 1 for
calculating the total volume of monomer solution
needed. Divide the total volume in half and prepare
this volume of each of the higher and lower %
acrylamide solutions.
5
Pour the “heavy” solution into the reservoir chamber
(the chamber furthest from the outlet). Open the stop-
cock long enough to displace air in the connection
between the chambers and then close. Fill the light
solution into the mixing chamber and place a stirring
bar in this chamber. Place the gradient maker on a
magnetic stirrer and begin stirring at a rate that does
not introduce bubbles into the solution.
•
p10
Note: 15% solid sucrose (w/v)
or 25% glycerol (w/v) in the
“heavy” solution minimizes
mixing and aids gradient
layering.