Hi All, Our protein is expressed as inclusion bodies ibn e.coli W3110 cells. we harvest the cells supend it in water [no buffer] and perform sonication to lyse the cells. we lyse the cells till our O.D is decreased to 1/3rd of original our starting O.D of suspension is about 250. then we centrifuge the abvoe suspencion at 12000 rpm to collect teh inclusion bodies. but we are not getting good yield and also inclusion bodies are not that pure. Do we need to add lysozyme prior to lysis. can anyone suggest a good method to lyse ecoli cells expressing protein in inclusion bodies.
Thanks in anticipation meg
