I express my protein in E. coli. After pelleting down the induced medium, I begin lysing the cell pellets through sonication. Suddenly, I am observing that my cells are not lysing. What could be the reasons? How can I fix this?
Dear Bonodip Chowdhury
to give you an opinion we need to have more information about:
- sonicator apparatus. sonicator tips size, number and lenght of pulses appled , amount of cell pellet that you lisate and volume and composition of the buffer used for cell resuspension. since many different aspect can affect the final result.
best
Manuele
Firstly, you need to make sure the sonication is working properly. Do you hear the big noise during the sonication process? If the sonication works well, then you may need to dilute the cell pellet/suspension with a bit more of the lysis buffer, to get a higher efficiency of lysis.
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