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Effect on PAGE gel

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Describe the effects the following would have on the way you PAGE gel would run:
a) Increasing the percent of acrylamide
b) Increasing the pH of the Tris-glycine buffer to 9.5
c) Decreasing the voltage at which the gel was run
d) Increasing the voltage at which the gel was run by a factor of 3
e) Doubling the sample load volume

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Solution Summary

This solution describes how a PAGE gel run would change under 5 different circumstances.

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Describe the effects the following would have on the way you PAGE gel would run:

a) Increasing the percent of acrylamide

Response: Acrylamide is the basic matrix material for PAGE. Therefore, if we increase the percentage of acrylamide, we will increase the amount of polymerizing within the matrix. This will make the gel physically "firmer." But more than that, the biological material running through the gel will be retarded more because they have to make their way between the acrylamide matrix. Therefore, there will be a greater resistance to the movement of biological molecules. Practically speaking, when we want to separate lower molecular weight material we increase the acrylamide concentration. This allows for the better separation of lower molecular weight species while tending to "jam together" and retard the migration of larger molecular weight material. Therefore, ...

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