In: Biology
For the gel electrophoresis system, what would be the consequence if the buffer added to the outer chamber was higher than the lip of the outer plate?
Electrophoretic systems are of two types. Vertical systems and horizontal system.
In horizontal gel electrophoresis, there is one chamber where buffer is filled and gel is kept in that buffer filled chamber. However in vertical electrophoresis system (generally used for SDS-PAGE), two chambers (Outer and inner chamber) are present. It is a general practice that in inner chamber is filled to the top however, in outer chamber lesser amount of buffer is filled.
Electrophoresis techniques operate on continuation of electric circuit. If the buffer added on outer chamber will be higher than the lip of outer plate then the circuit will be broken, Therefore, the material (Protein) will not be separated/resolved.
In continuation of this error if the level of buffer reaches further above on the top of the plate then, the circuit will be reversed and the whole loaded sample can be ejected in the buffer.
Vertical gel electrophoretic techniques like SDS gel electrophoresis plays vital role in separating different proteins. Proteins are separated on the basis of their molecular weight. The molecules are separated by directional flow of charged proteins or other molecules in of buffer, Buffer plays a key role in maintaining the electric circuit. If buffer is in very little amount, even then the circuit is broken and protein fails to separate or resolve.