Demonstration of Size-Based Separation of Molecules by Gel Chromatography: An Exercise for Biology Beginners

An introductory laboratory exercise has been designed for biology beginners to visualize and analyze the chromatographic separation of a mixture of blue dextran, α-chymotrypsinogen (protein) and potassium ferricyanide on a Sephadex G-75 column (601.0 cm). Separation of the two coloured components i.e. blue dextran (blue colour), α-chymotrypsinogen (colourless) and potassium ferricyanide (yellow colour) of a green-coloured mixture can be visually seen in the form of blueand yellow-coloured bands distant by a colourless zone. The elution volumes of different components in the mixture were found similar to the elution volumes of these components, when loaded individually onto the same column. Such demonstration of separation of different components in a mixture on a gel chromatographic column is an interesting exercise for biology beginners (undergraduate students) to learn separation technique on the basis of size. [Tan CY, Tayyab S. Demonstration of Size-Based Separation of Molecules by Gel Chromatography: An Exercise for Biology Beginners. Life Sci J 2012;9(4):1560-1563] (ISSN:1097-8135). http://www.lifesciencesite.com. 237