Principles of Biochemistry (2+1)

Procedure

• Fix the column vertically to a stand.
• Equilibrate the column with 4ml Gel filtration buffer. Drain out the buffer completely.
• Load 0.2ml of the sample onto the column, along the sides of the column.
• Allow the sample to sink completely.
• Allow the buffer to flow out completely.
• Keep filling the column with buffer, till all the coloured biomolecules have eluted out (approx. 20 ml buffer is needed).
• Collect the coloured fractions in different tubes.
• Fix the bottom cap and then the top cap to stop the flow of buffer.

                                        Observation     

• Note down the time and order in which various molecules leave the column.

       (Store the column at 4°C for next use.)