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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