japonet
What is S japonicus

 

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Media

We use general S. pombe media for culturing. Although there are several differences in character (iodine negative, eight spores or lithium super sensitivity), we do not have much difficulty in manipulating them. For solid media, we use 20g/L (YE) and 24g/L (EMM2). NOTE: WAKO agar #010-15815 is used or EMM2 media.

Growing media

YE    

 5 g/L         yeast extract,
30 g/L       D-glucose
Culturing Media (for selection or sporulation)
EMM2

2.2 g         Na2HPO4,
3.0 g         potassium hydrogen phthalate,
5.0 g         NH4Cl,
20 g    D-glucose,
1x      salt stock,
1x         vitamin stock,
1x         mineral stock,
(50x salt        53.5 g MgCl2.6H20, 0.74 g CaCl2.2H2O, 50 g KCl and 2 g Na2SO4)
(1000x vitamin stock/L: 1 g sodium pantothenate, 10 g nicotinic acid, 1 g inositol and 10 mg biotin)
(10,000x minerals: 5 g H3BO3, 4 g MnSO4, 4 g ZnSO4.7H2O, 2 g FeCl3.6H2O, 0.4 g H2MoO4.H2O, 1.0 g KI, 0.4 g CuSO4.5H2O and 10 g citric acid)
supplemented with 50 mg/L of amino-acids or nucleotides
NOTE: WAKO agar #010-15815 is used or EMM2 media.
YE+FOA YE media supplemented with 50 mg/L uracil and adenine. 1.5 mg/ml 5-FOA was used.
YE+Geneticin 40 mg/L Geneticin is used. For selecting more than 108 cells on 90 mm plates, at least 40 mg/L of Geneticin is required. However, for normal use, 20 mg/L Geneticin is enough to kill S. japonicus cells. Currently we have been using one from ALEXIS (380-013).
YE+clonNAT 100 mg/L is used. The ClonNAT gives less false positive than Geneticin.
SPA   10 g       glucose,
1 g         KH2PO4,
30 g         agar,
　　　　supplemented with 100 mg/L of adenine, arginine, uracil and leucine
Reference; Moreno, S., Klar, A. and Nurse, P. (1991). Molecular genetic analysis of the fission yeast Schizosaccharomyces pombe. Methods Enzymol. 194:795-823.

Microbial Physiology Laboratory (Niki Laboratory)
Department of Gene Function and Phenomics, National Institute of Genetics

1111-1 Yata Mishima Shizuoka Japan P.O. 411-8540