Dear all,
I have been trying to express a recombinant protein which is almost 86 kDa in Pichia pastoris. My transformation into pichia genome was successful ,as confirmed by colony PCR, but I am having a problem in expression. I maintain all the parameters suggested in Pichia expression system. I know that Oxygen is a vital parameter for pichia to grow and therefore should I not use aluminium foil to cover the conical flask? or should I cotton plug it? I was advised to use a small beaker to cover the mouth of the conical flask,thus allowing proper oxygen. What is the ideal way to use a conical flask system for pichia pastoris expression system?
Hi!! I used to cover my flask with three layers of cheesecloth and secure with a rubber band, it worked perfectly. Cheers.
Hi. Try to use big conical flask with less culture volume, so the aeration will be optimal.
Hello. To cover the flasks, you can use few layers of sterile gauzes or porous tapes. In order to better aeration, It is recommended that the volume ratio of the culture medium to the flask should be between 10-20% and shaking at 225-300 rpm to allow air to flow well into the flasks.
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