Extreme media preparation experiment

Summary

Extreme media are media that contain only carbohydrates and inorganic salts that provide nitrogen, phosphorus, and trace elements. Source of content: compact guide to molecular biology 5th ed.

Operation method

Extreme media preparation experiment

Materials and Instruments

Water Na2HPO4 KH2PO4 NH4Cl Antibiotics
Glass bottles Flasks

Move

1. In a 2L flask, add the ingredients of the following formula to water and heat and stir until dissolved.

(1) 5 X M9 medium, per liter

30 g _Na2HPO4

15 g _KH2PO4

5 g _NH4Cl

2.5 g NaCl

15 mg _CaCl2 (may or may not be added)
(2) 5 X M63 medium per liter

10 g ( _NH4)2SO4

68 g _KH2PO4

2.5 mg _FeSO4 - _7H2O

pH adjusted to 7.0 with KOH (3) 5 X A medium, per liter

5 g ( _NH4)2SO4

20.5 g _KH2PO4

52. 5 g <sub>K2HPO4

2.5 g citric acid dihydrate</sub>

(4) 1 X medium

Dilute 5X medium with sterile water and add the following sterile solution per liter

1 ml 1 mol/L _MgSO4 - _7H20

10 ml 20% carbon source (sugar or glycerol)
2. Pour into glass vials, loosen the lids and ^autoclave at 15 ^lb/in2 for 15 min.

3. Allow the medium to cool to below 50°C (at this temperature the bottle may feel hot when held but can be held), add the other supplements and antibiotics, and cool the glass bottle to below 40°C with the lid tightly closed.

4. May be stored indefinitely at room temperature.

Common Problems

1. The following solutions may also be added if required

0. 1 ml 0.5% vitamin _B1 (thiamine)

5 ml 20% casein hydrolysate (casamino acid) (Difco) or L-amino acids to 40 μg/ml or DL-amino acids to 80 μg/ml

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