Hi All,
We are interested in using the Branson SFX150 sonicator we have in our lab to lyse DE3 E.coli in order to purify a couple of proteins. I have worked with larger sonicators in the past to this end, but I was wondering if anyone out there has used one of these smaller wattage units to successfully sonicate ~1L cultures of bacteria.
I know we will need the 1/4" probe available for the unit in order to process samples between 10-100mL (ours will most be 1L culture worth of pellet resuspended in 30-50 mL range ).
Does anyone have any suggestions on amplitude values and pulse times that have worked for them in the past?
Thank you!
Dear Alex
generally in my experience volumes from 30 to 100ml can be sonicate with standard probe, 40% amplitude, 30 second on and at least 2' minutes off to Cool down the sample for 10 cicles. Put the sample in a Plastic Becker and keept on ice during sonication.
good luck
Manuele
My advice would be to check for lysis by taking appropriately diluted samples at different times of sonication and following the drop of OD600. Unless your cells contain inclusion bodies (check by inspecting them under a phase contrast microscope), you should achieve a > 70% drop relative to the unprocessed sample. Be careful not to trap air below the sonicator probe to avoid foaming. But personally, if available, I would prefer using a French press rather sonication.
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