Medium data

Search for medium no. = [499].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.


Solution A:
Mineral solution (see below)  50.0     ml
Trace element solution SL-10 (see Medium No. 433)  1.0     ml
Rumen fluid, clarified (see Medium No. 266)  50.0     ml
Trypticase peptone (BD-BBL)  1.0     g
Vitamin solution (see below)  5.0     ml
Crotonic acid  1.7     g
Resazurin  1.0     mg
Distilled water  810.0     ml
Solution B:
NaHCO3  3.5     g
Distilled water  70.0     ml
Solution C:
L-Cysteine·HCl·H2O  0.3     g
Distilled water  10.0     ml
Solution D:
Na2S·9H2O  0.3     g
Distilled water  10.0     ml
Mix ingredients of Solution A, adjust pH to 7.2, bring to a boil for a few minutes and cool down to room temperature under a stream of N2-CO2 (4:1, v/v) gas mixture. Distribute the solution into culture vessels under the same gas mixture, seal with butyl rubber stoppers and autoclave. Filter-sterilize Solution B and equilibrate by gassing with the same gas mixture for at least 15 min. Autoclave Solutions C and D under a N2 atmosphere. Add appropriate amounts of Solutions B, C and D to Solution A to complete the medium.
Mineral solution:
KH2PO4  10.0     g
MgCl2·6H2O  6.6     g
NaCl  8.0     g
NH4Cl  8.0     g
CaCl2·2H2O  1.0     g
Distilled water  1.0     L
Vitamin solution:
Biotin  0.25    mg
Nicotinic acid  2.5     mg
Thiamine·HCl  1.25    mg
p-Aminobenzoic acid  1.25    mg
Pantothenic acid  0.62    mg
Pyridoxine·HCl  6.2     mg
Distilled water  1.0     L