“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.