Below is an example of what can happen to a gel with a bad buffer.
The pH of this buffer is the likely suspect. After testing the pH
meter to ensure its accuracy and mixing up new TBE running buffer, a great
improvement was seen.
Before: Notice particularly the bands found in the lanes containing
the ladders with known marker weights/sizes. These are labeled 25bpo,
25bp, and 1Kb. At each site where there ought to be 1 band there
are 3-4 bands in the 25bp lanes and 2 bands in the 1Kb ladder lane.
After: Here the ladders are back to normal, with single bands
at each location. (However, the gel still has a slight "smile" to
it, since nothing was used to evenly disperse heat generated during the
run across the gel. For even greater improvement see the page describing
the use of copper plates.)
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