Gel Electrophoresis

  1. Make 1x tris-glycine-SDS running buffer. (For a bis-tris gel, use MOPS-SDS running buffer).
  2. Load the gels into the gel holders being sure to keep a tight seal between the gel- cast and the gasket.
  3. Pour the running buffer into the middle of the gels and check for leaks.
  4. Pour the rest of the running buffer into the bottom of the running container.
  5. Remove combs and use a pipette to clean away any unpolymerized acrylamide.
  6. Make a marker for each gel (10 microliters marker + 10 ul blue SDS-buffer)
  7. Load all 20 ul of the marker into the first well.
  8. Load 25 ul of the lysate (1 ug/ul) into the rest of the wells.
  9. Fill any empty wells with Laemmli SDS-buffer.
  10. Set gel to 90v and run for approximately 2 hours.

 

 

Leave a Reply