Technical Support FAQ

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Technical Support FAQ


Question:

The bands on my gels are misshapen, skewed, or distorted, instead of flat, thin lines. What can I do?


Answer:

1) There may be poor polymerization around the wells. Avoid polymerizing the stacking gel with bubbles under any teeth of the comb. Degas the stacking gel solution thoroughly with a bench vacuum or better before casting the gel. The vacuum obtained by an aspirator on the sink is not adequate in most cases. (See Bulletin 1156 for information on polymerization of acrylamide.) Increase the concentration of APS (1610700) and TEMED (1610800) by 25%. Just before casting the stacking gel, wipe the comb teeth with TEMED.

2) Distorted bands can be caused by too much salt (too high ionic strength) in the sample. Remove extra small ions by dialysis or desalting column. Both serum and ascites fluid, for example, must be desalted before adding the sample buffer.

3) The gel interface could be distorted due to polymerization occurring too fast. Decrease the polymerization rate by descreasing the amount of TEMED by 25%. Overlay the gels very carefully.

4) The wells have not been rinsed out before addition of the sample. Unpolymerized acrylamide in the wells can cause distorted bands. Rinse the wells out thoroughly before addition of the sample.


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