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S L Forsburg nospamforsburg at salk.edu
Tue Apr 13 16:36:29 EST 1999

> Endoreplication in S. pombe ?
> Steve K. Cho (cho2 at simmons.swmed.edu)
> Mon, 12 Apr 1999 12:19:35 -0600
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> Message-Id: <199904122000.NAA15320 at net.bio.net>
> To: yeast at net.bio.net
> Date: Mon, 12 Apr 1999 12:19:35 -0600
> From: "Steve K. Cho" <cho2 at simmons.swmed.edu>
> Subject: Endoreplication in S. pombe ?
> Dear members of yeast community,
> I would like to know more about endoreplication in S. pombe that produce
> homozygous diploids.
> If anyone familiar with this topic, help me to learn more about
> endoreplication.
> How can I obtain those homozygous diploid strains?

Easy to do, using the indicator phloxin B.  Diploid cells do not 
exclude this vital stain as easily as haploids, and therefore form 
darker pink colonies on plates.  Simply plate a strain of interest 
to single colonies on YE plates with phloxin B.  Pick any dark pink
colonies and restreak.  Analyze under the microscope for diploid 
morphology (diploids are longer and wider than haploids) and best 
to also double check ploidy by FACS.

Endoreduplication rates vary in different strains 
 but in my experience you can pick up diploids at between 
1 in 10^3 or 10^4.

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S L Forsburg, PhD  forsburg at salk.edu
Molecular Biology and Virology Lab          
The Salk Institute, La Jolla CA 

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