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Gel Electrophoresis - Separation Of Proteins

size dna concentration pores

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Proteins are usually separated using vertical polyacrylamide gel electrophoresis (PAGE), a process that separates them on the basis of their electric charge and their size. Proteins with a greater negative charge will be attracted more strongly and move faster toward the anode. The charge density on the proteins would cause smaller molecules to move more quickly through the gel's pores.

The size of a gel's pores can be changed depending on the size range of the proteins being separated. This is done by raising or lowering the concentration (1) DNA fragments are loaded into the gel. (2) Electric current is turned on, pulling the negatively charged DNA toward the positive electrode. (3) Smaller DNA fragments move faster through the gel. (4) The DNA is stained, revealing its position. of acrylamide and bisacrylamide in the gel. Increasing the concentration results in more crosslinking between the two components, decreasing the pore size. Decreasing the concentration increases the pore size of the gel. Small proteins are separated better in a gel with large pores.

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