“M” or minimal media (Sager & Granick)
(low ionic strength medium to decrease palmelloids and encourage flagella growth)

Solution #                                       For 1 Liter

1.    10X Trace metal stock            1.0 ml
2.    10% Na Citrate•2H20              5.0 ml
3.    1% FeCl3•6H2O                        1.0 ml
4.    5.3% CaCl2•2H2O                    1.0 ml
5.    10% MgSO4•7H2O                  3.0 ml
6.    10% NH4NO3                           3.0 ml
7.    10% KH2PO4                            0.7 ml
8.    10% K2HPO4•3H2O                1.5 ml (start with 1.2 ml, use to pH)
Add components to distilled water in the order given
pH 6.8

10X Trace Metal Stock Solution

Component                           mg/L in 10X stock

1.    H3BO3                             1000
2.    ZnSO4•7H2O                  1000
3.    MnSO4•H2O                    303
4.    CoCl2•6H2O                     200
5.    Na2MoO4•2H2O             200
6.    CuSO4•5H2O                   70

“M-N” medium (for gametogenesis)
Same as M but omit solutions 6 and 7 and double the amount of 8.  In some cases, better mating is observed when gametes are prepared in M-N + 10mM HEPES, pH 7.0.

“N” medium (nitrate as nitrogen source)
Omit solution 6 and add 4 ml 1M KNO3

“R” medium (acetate as carbon source)
Same as M but add 10 ml of 2.2M NaAc/Liter after all of the components are added.  Also, increase solution 7 and 8 by 3X.

“½ R” medium (sometimes used for mating plates to improve survival of tetrad progeny)
Same as M but add 5 ml of 2.2M NaAc/Liter after all of the components are added.  Also, increase solutions 7 and 8 by 3X.

Arginine medium (used for arginine auxotrophs)
Same as M but add 0.5 ml of 10% of L-arginine/Liter of media before autoclaving.

Arginine medium for mating plates (to increase survival of arg- progeny)
Same as M but add 2 ml of 10% of L-arginine/Liter of media and reduce solution 6 to 0.3 ml/L.

SGII medium (used for transformation with glass beads)
Use solutions 1 – 6 as for M medium, then add:
2 g/L         NaOAc•3H2O
3.67 g/L    NaH2PO4•H2O
1.5 g/L       K2HPO4
pH 6.2

SGII/NO3 medium (used for selection of NIT+ transformants)
Make SGII as above, but omit solution 6 and add 4 ml 1M KNO3.

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