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S.A.P's Content

There have been 3 items by S.A.P (Search limited from 22-September 19)

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#88960 Pichia transformant selection

Posted by S.A.P on 07 October 2010 - 02:22 AM in Molecular Biology

Can anyone tell me why sorbitol is used during pichia transformant selction? All I've read about it is that it hinders growth etc!

#60888 Sonication issues!

Posted by S.A.P on 02 March 2010 - 02:49 AM in Protein Expression and Purification

I always sonicate at a 1:3 ratio (5 mins actual sonication time, 15mins off) per pellet from 200ml of culture. My samples are always kept on ice during the procedure too. The amplification was 40%.
I was trying two different buffers, one was something I have always used (tris, DTT, EDTA and NaCl at pH 8), the other was bought from Qiagen. But I didn't get a pellet for either once I had spun them down!

I've never had this problem before but I guess I will soon find out if this really matters soon enough!

#60794 Sonication issues!

Posted by S.A.P on 01 March 2010 - 04:46 AM in Protein Expression and Purification

Following sonciation of my cells, I centrifuged as normal to harvest the insoluble protein fraction. Only to find I couldn't get a significant pellet no matter how much I spun the samples down!

Any ideas? Perhaps the sonication time was excessive?

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