350
w. B. REDMOND et al.
FeC13.6HzO
Agar (certified)
Distilled water
1.0 mg
485 ml
11.0 g
Melt agar; add MgS04 and CaC12. Autoclave at 15 lb pressure for 20 min.
Add 1 ml of Fe3 stock solution (page 352).
Solution D, Sodium glutamate (stock solution) (page 352), 5 ml.
Solution E, Oleic acid-albumin complex.
Albumin (bovine Fraction V)
3.5 g
Distilled water
95 ml
Oleic acid-NaOH mixture?
5 ml
NaOH (2.5 N)
0.35 ml
Sterilise by Seitz filtration. Warm to about 45°C.
Mix solutions A, B, C, and D after each has been sterilised as indicated.
Allow to cool to 48°C and add Solution E. Mix gently so as to prevent the
formation of bubbles. Pour into Petri dishes, 30 ml/plate.
Allow the medium to solidify. Dry in incubator for 48 h. Store inverted
in refrigerator. Do not use when more than three weeks old.
B. RVB-10 liquid medium. Nutrient broth
A liquid medium without agar similar to the above has been found to
produce good growth of mycobacteria for phage studies. Tween-80 is
added to the RVB medium in order to produce dispersed growth of some
mycobacteria.
RVB-10
Solution A
NazHP04 (anhydrous)
KH2P04
NaCl
NH4Cl
Sodium pyruvate
Nutrient broth
Casein hydrolysate (enzymatic)
Glycerol
ZnS04
C U S O ~ .
5Hz0
Tween-80 (stock solution) (page 352)
Distilled water
t Add 0.12 ml oleic acid to 10 ml 0.05 N NaOH.
1.2 g
1.0 g
2.5 g
0-5 g
0.75 g
4.0 g
2.0 g
8.0 ml
0.10 mg
0-10 mg
2 ml
360 ml