Medium data

Search for medium no. = [574].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

574   DB CHARACTERIZATION MEDIUM NO. 2

MDS salt water (see below)  833.0     ml
FeCl2 solution (see Medium No. 187)  1.0     ml
Trace element solution (see Medium No. 187)  1.0     ml
Peptone (Oxioid)  0.25    g
Yeast extract (BD-Difco)  0.05    g
Add components and bring volume to 980 ml with distilled water. For preparation of solid medium, add 20 g/L agar (if necessary, use washed agar as described below). Autoclave, and after cooling, add the following solutions (filter-sterilized or *autoclaved) to the medium:
1 M NH4Cl  5.0     ml
Potassium phosphate buffer* (see below)  2.0     ml
Vitamin solution (see below)  3.0     ml
1 M Sodium pyruvate solution  10.0     ml
Readjust pH to 7.0 - 7.5 with a sterilized 10% Na2CO3 solution, if necessary.
Washed agar:
Stir agar in distilled water and stand still for 30 min. Then, remove supernatant by decantation. Repeat the washing at least twice.
MDS salt water:
NaCl  240.0     g
MgCl2·6H2O  30.0     g
MgSO4·7H2O  35.0     g
KCl  7.0     g
1 M CaCl2 solution  5.0     ml
Add components to distilled water and bring volume to 1.0 L. Adjust pH to 7.5 with 1 M Tris base.
Potassium phosphate buffer:
1 M K2HPO4  83.4     ml
1 M KH2PO4  16.6     ml
Combine the two solutions and check that pH is close to 7.5. Add an equal volume of distilled water.
Vitamin solution:
Biotin  3.0     mg
Folic acid  4.0     mg
Pyridoxine·HCl  50.0     mg
Thiamine·HCl  33.0     mg
Riboflavin  10.0     mg
Nicotinic acid  33.0     mg
DL-Calcium pantothenate  17.0     mg
Vitamin B12  17.0     mg
p-Aminobenzoic acid  13.0     mg
Lipoic acid  10.0     mg
Distilled water  1.0     L