To prepare the wells, follow these steps:
Action
Step
De-aerate the stacking gel monomer solution.
1
Add catalyst and initiator to the stacking gel monomer solution and then
pour.
2
Use a pipette to deliver the solution into one corner of the plate, taking care
not to trap any bubbles.
3
Insert a comb (at a slight angle to prevent trapping air) into the sandwich,
allowing the comb sides to rest on the spacers.
4
Overlay each gel with a thin layer of water-saturated n-butanol, water, or
diluted gel buffer to prevent gel exposure to oxygen. Slowly deliver the
overlay solution from a glass syringe fitted with a 22-gauge needle. Apply
the solution near the spacer at the side of the sandwich and allow it to flow
across the surface unaided.
5
Allow a minimum of one hour for the gel to polymerize.
6
To prepare the sample, follow these steps:
Action
Step
Increase liquid sample density with 10% glycerol or sucrose.
1
Add a tracking dye.
2
For SDS protein gels, use 2× treatment buffer to denature both liquid and
dry samples in a test tube.
3
To liquid protein solutions, add an equal volume of 2× buffer.
To dry protein samples, add equal volumes of buffer and ddH
2
O to achieve
the desired concentration.
Heat the tube in boiling water for 90 seconds, then chill it in ice until ready
to use.
4
Treated samples can be stored at -40°C to -80°C for future runs.
To load samples, follow these steps:
SE 250/260 Mighty Small II Operating Instructions 29281629 AA
37
5 Operation
5.2 Assembly
