Phloxin B is a stain that accumulates in dead cells, which become stained dark red. This can be confirmed by microscopic examination of the cells. Streaking of strains on a plate next to a known haploid can help identification by having cells of both types in the same microscopic field. Often sectored colonies are seen. PCR can also be used.
Temperature is quite critical.
Gel purified band can be directly sequenced. Analyze in the agarose gel. The auxotrophic markers most commonly used in S. pombe require adenine, glutamic acid, histidine, leucine, lysine and uracil, although others are available. To test for auxotrophy the strain is grown up to single colonies on YES and then replica plated to minimal medium with and without the appropriate supplement. The plates are incubated for 1 - 2 days and then examined for growth under these conditions.