What's the criteria of comolete lysis of E.coli when breaking them using ultrasonic? What phenomena can help to determine if the cell are broken completely? From smell, transparence of the lysis or SDS-PAGE of the cell pellet and solution?
After breaking them, normally you centrifuge at like 3000 xg, to pellet unbroken bacteria and debris. Check your pellet (and its size) after centrifugation if there are any bacteria left (size of pellet or microscopy).
Jorgaq Pata thanks! Pellet would be bigger if bacteria are not broken? I normally spin at 18000 rpm for 20 min.
Xiang Zhang well yes, normally you have debris that are pelleted too, but if a lot of bacteria are unbroken, the pellet will be bigger.
What i did once, is that i sonicated several times, and i observed the lysate in microscope. It was nice and conclusive.
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