Hello!
My Pichia pastoris cells were lysed using Yeast Buster and then precipitated using the methanol/chloroform method. The pellet was a circular flat white layer (like a membrane) and I couldn't resuspend it in my 8M Urea % CHAPS Lysis Buffer.
In another attempt, I tried grinding the cells in liquid nitrogen, and directly precipitate it in TCA/acetone. The pellet was unusually a lot, and similarly, very hard to resuspend in the Lysis Buffer.
Has anyone experienced this before? I would like to know what could be in that pellet that makes it so difficult to resuspend.
Thank you!
Ally
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