Materials and Reagents

1. Yeast strain (Schizosaccharomyces pombe)
   
2. Flask (IWAKI PUMPS, catalog number: 4980FK500 )
   
3. Conical tube (BD Biosciences, Falcon^®, catalog number: 2070 )
   
4. 1.5 ml microcentrifuge tube (Eppendorf, catalog number: 022364111 )
   
5. Lysing enzymes from Trichoderma harzianum (Sigma-Aldrih, catalog number: L1412 )
   
6. Zymolyase 100T (Seikagaku Corporation, catalog number: 120493-1 )
   
7. Complete Mini (Roche Diagnostics, catalog number: 11836153001 )
   
8. Anti-GFP antibody (Roche Diagnostics, catalog number: 11814460001 )
   
9. Anti-histone H3 antibody (Millipore, Upstate Biotechnology, catalog number: 05-499 )
   
10. Anti-alpha-tubulin antibody (generously provided by Dr. A. Baines)
    Note: Commercially available antibodies against alpha-tubulin (such as Abcam, catalog number: ab6161 ) can be used.
    
11. Goat HRP conjugated-anti-mouse antibody (Life Technologies, Biosource, catalog number: A10551 )
    
12. YES liquid medium (see Recipes)
    
13. STOP buffer (see Recipes)
    
14. PEMS buffer (see Recipes)
    
15. 2x HBS buffer (see Recipes)
    
16. Lysis buffer (see Recipes)
    
17. 2x Laemmli protein sample buffer (see Recipes)
    

Equipment

1. Air- (Tykyo Rikakikai, Eyela, model: FMC-1000 ) or water bath- (Taitec, model: MM-10 ) incubator shaker
   
2. Centrifuges equipped with 50 ml tubes (Tomy Digital Biology, model: AX-501 ) and 1.5 ml microtubes (Tomy Digital Biology, model: MX-107 )
   
3. Heat block (TAITEC, model: DTU-1BN) or water bath (Taitec, model: EXN-B )
   
4. Light microscope (Nikon Corporation, model: Eclipse E200 )
   
5. Electrophoresis apparatus (Bio-Rad Laboratories, catalog numbers: 165-8002JA and 164-5052 )
   
6. Transfer unit (Bio-Rad Laboratories, catalog numbers: 170-3930JA and 170-3935JA )
   

Procedure