Gel electrophoresis is a technique
used for the separation of nucleic acids and proteins. Separation of large
molecules depends upon charge and mass.
When protein or DNA is mixed
in a buffer solution and applied to a gel, these two forces act together.
An electrical current is applied, so that one electrode repels the molecules
while the other electrode attracts the molecules. Smaller molecules
move further, while larger ones encounter difficulty in moving through
the gel and don't go as far.
During electrophoresis, macromolecules
are forced to move through the pores when the electrical current is applied.
Their rate of migration through the electric field depends on the strength
of the field, size and shape of the molecules, relative hydrophobicity
of the samples, and on the ionic strength and temperature of the buffer
in which the molecules are moving.
The result is stained so that the
separated macromolecules in each lane can be seen in a series of bands
spread from one end of the gel to the other. |
1. Prepare the
gel and place it in the chamber.