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- The height of the buffer in the chamber. Though you are putting in a specific voltage and amperage more buffer will increase resistance, and therefore take away from the speed at which your bands run. Looking from the side of the box 0.5-1.0 cm above the cell is usually what I run. Not more. Plus, try to keep it consistent from gel-to-gel.
- The initial run speed. Most people ignore this, but it's a very useful trick. If I'm running a 120V gel, I always run it at 90V until the sample is all the way out of the well and into the lane (this is usually ~10 minutes). This way I can be more confident that my samples are running at the same velocity through the well when it counts. Basically, this is allowing your sample more time to snake its way into the pores of the gel, rather than cramming it in.
- Clean the electrodes in the box (that interface with buffer) with some diluted acetic acid or soap.
- This one is obvious, but make sure you're using 1X buffer.
#147379 Plasmides run higher than marker
Posted
on 03 January 2013 - 09:04 AM
A couple of things to keep in mind:
