I am preparing a sonicate to destroy E.Coli CD41 DE3 cells. However, the sonicate efficiency does not seem to be high.
I follow the following procedure: Dry biomass is mixed with 1.5 ml of lysis buffer and then sonicated with the cycle: beat for 15 seconds, rest for 2 minutes and repeat 6 times.
I suspect due to suboptimal amount of wet biomass. Do you have any experience when choosing the amount of wet biomass to take away the sonicate?
First, your assumptions are correct, you have to take care of the amount of wet biomass. I don't know what concentration % you used, but I would recommend up to a maximum of 20%, mainly because it is a standard, but you need to make the necessary calculations specific to your substance, because of the energy requirements and water loss. In short, 15-20%.
Thank you Luis Alberto Manzo Navarro for answering my question. I tried again with 50mg wet mass per 1.5ml lysate and the results were better. After SDS-Page electrophoresis showed significant soluble protein. However, it seems that this ratio is higher than your suggestion. I will try with 15-20% to see the results.
I have calculated them to a total energy of 400J.
However, do you think the sonicate cycle should change?
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