Cell disruption liquid extrusion

Disrupt cells by liquid extrusion in four passes through liquid duidizer (Microfluidic Corp.) at 70 psi, bring the lysate volume to 800 ml. 

Resuspend 50 g of Pyrococcus furiosus cells in 200 ml of buffer A. Disrupt cells by liquid extrusion in three passes through liquid fluidizer (Microfluidic Corp.) at 70 psig. Immediately after lysis add Triton X-100 to cell lysate (250 ml) to a final concentration of 0.1% (w/v). Clarify cell lysate by centrifugation in Sorval GSA rotor at 11,000 rpm for 1 hr. Add 20 ml of 5% PEI (titrated to pH 7.5 with... 

Homogenize cell pellet in 150 ml of buffer A and disrupt cells by liquid extrusion in three passes (see procedure for 5. acidocaldarius above). 

Liquid shearing, such as high pressure homogenization, solid shear, such as bead milling and the extrusion of frozen cells are the most common mechanical methods for cell disruption (D souza and Killedar, 2008 Hatti-Kaul and Mattiasson, 2003). These methods are mainly based on the use of mechanical force to disrupt the cell. Some of these techniques, such as bead milling, have already found applications on a large scale (Prasad, 2010). 

---