gene knock-out in pombe

Wen Song wensong at uclink.berkeley.edu
Tue Jan 27 13:34:18 EST 2004


I am trying to knock-out a gene in pombe. However, I don't have enough experiecne to work with 
diploid. Thanks a lot for Forsburg's pombeweb, I learned a lot of from there. However, I still 
have some questions.

1. What is a good diploid to use for gene knock-out?
2. Can I avoid doing tetrad analysis (there is no equipment or technical know-how around) even if 
my gene is essential? Can I just do a radom spores and check for my selection marker, say Ura4, to 
see if my gene is essential?
3. Is there a detailed protocol I can follow?

I really appreciate your input and advics.

Thanks a lot in advance!

Wen Song


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