Comparison of lysis methods Sonication
- most efficient method of cell fractionation
- problem: heat build up which can denature proteins (proportionate to length of sonication)
- precaution: do on ice and sonicate intermittantly
material: ultrasonication bath or rods
Homogenisation
- best for animal tissue; less suitable for cells
- precaution: do on ice to reduce heat build-up and denaturation
material: drills (polytron), pestle/tube (dounce) bead beaters
Freeze-thaw
- least effective method
- plus: does not denature proteins as much as other methods
material: pestle and mortar, liquid nitrogen
Detergents
- chemical method of lysis
- problems: detergent may inhibit subsequent reactions
- problems: detergent may disrupts protein interactions
no additional material required, just the chemicals and typical tubes
