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Main sol. 9a

Internal Solution-ID: S10

Equipment needed: Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Baker's yeast 2.50 g 2.5 -
CaCl2 x 2 H2O 1.36 g 1.36 9.251
Vitamin B12 0.50 mg 5.0e-4 3.7e-4
Agar
(Difco)
8.00 g 8 23.786
Distilled water 1000.00 ml - -
1 Sterilize vitamin B12 separately by filtration. Prepare and store yeast cells as autoclaved stock suspension (5 g baker's yeast/100 ml distilled water, adjust pH to 6.5 and autoclave).
2 Adjust pH of medium to 7.2 with KOH before as well as after autoclaving and cooling to 50°C (use pH-indicator paper).
3 For suspension of freeze-dried cells from ampoules add about 0.5 - 1.0 ml medium MD1 (per liter: casiton, 3.0 g; CaCl2 x 2 H2O, 0.7 g; MgSO4 x 7 H2O, 2.0 g) to the vial with freeze dried material.
9a VY/2, REDUCED MEDIUM