I try to express some protein in Pichia pastoris.
And I have the problem: when I carry out electroporation and spread yeast on YPD plates with zeocine ( 100ug/ml, 500ug/ml ), yeast grow only one day and the plate is full of yeast no colonies just coating. I tried to spread P.pastoris without vector contained zeocine resistence ( pPICZalpha)like negativ control. And the plate is coating again. The same I have when I spread E.coli nova blue or TOP10 on the LB low salt medium plates containing zeocine. I changed zeocine,
I controled preparing of medium and the result is same.
I do not know what I do now.
Could you help me somebody?
Thank you very much
Edited by sius, 24 August 2010 - 04:40 AM.