ABSTRACT

It is not an exaggeration to state that now the most common method of cell breakage for the isolation of intact walls, is to shake them in the presence of glass beads of a diameter around 1 mm. The types of instruments used depends on the size of the sample, but may go from the most sophisticated ones, to the use of simple centrifuge tubes agitated in a vortex-type mixer. Taking into consideration that the cell wall contains proteins, and that proteases are liberated during the breakage procedure, it is common to add protease inhibitors to preserve them. Depending on the future analysis to be made, the walls are washed with buffer and saline solutions, and if necessary, with detergents or organic solvents. Purity of the isolated walls is then normally con…rmed by microscopic analysis (either light or electronic, as required). A thorough review of all these procedures was described in the …rst edition of the book (Ruiz-Herrera 1992).