BAM Media M200: Selective Middlebrook 7H11 Agar
October 2023
|
Middlebrook 7H11 Agar base * |
20.5 g |
|---|---|
|
Middlebrook OADC Enrichment ** |
100 ml |
|
Glycerol |
5 ml |
|
Distilled water |
800 ml |
|
Trimethoprim Lactate stock solution |
50 ml |
|
Amphotericin B stock solution |
50 ml |
|
Polymyxin B stock solution |
1 ml |
|
Carbenicillin stock solution |
1 ml |
|
Middlebrook 7H11 Agar base * |
20.5 g |
|---|---|
|
Casein enzymatic hydrolysate |
1.00 g |
|
Ammonium sulfate |
0.50 g |
|
Monopotassium phosphate |
1.50 g |
|
Disodium phosphate |
1.50 g |
|
Sodium citrate |
0.40 g |
|
Magnesium sulfate |
0.05 g |
|
L-Glutamic acid |
0.50 g |
|
Ferric ammonium citrate |
0.04 g |
|
Pyridoxine |
0.001 g |
|
Biotin |
0.0005 g |
|
Malachite green |
0.001 g |
|
Agar |
15.00 g |
|
Middlebrook OADC Enrichment ** |
100 ml |
|---|---|
|
Bovine Albumin Fraction V |
5 g |
|
Dextrose |
2 g |
|
Catalase |
4 mg |
|
Oleic acid |
50 mg |
|
Sodium chloride |
0.85 g |
|
Distilled water |
100 ml |
* Middlebrook 7H11 Agar base are available from Fisher Scientific (Catalog No. R454002), MilliporeSigma (SKU M0428), and other vendors with equivalent formulations.
** Middlebrook OADC Enrichment are available from Fisher Scientific (Catalog No. R450605), MilliporeSigma (SKU M0678), and other vendors with equivalent formulations.
Suspend 20.5 g of Middlebrook 7H11 Agar base in 800 ml of Distilled water containing 5 ml of Glycerol. Heat to boiling to dissolve completely. Sterilize by autoclaving. Cool to 45-50oC and aseptically add 100 ml of Middlebrook OADC Enrichment, 50 ml of Trimethoprim Lactate stock solution, 50 ml of Amphotericin B stock solution, 1 ml of Polymyxin B stock solution, 1 ml of Carbenicillin stock solution. Mix and dispense. Final pH 6.6 +/- 0.2 at 25oC. Store the plates at 2-8oC with expiration in 90 days.
Trimethoprim Lactate stock solution. Dissolve 100 mg of Trimethoprim Lactate in 250 ml of Distilled water completely. Filter-sterilize the solution and store at 2-8oC with expiration in 183 days.
Amphotericin B stock solution. Adjust 260 ml of Distilled water with NaOH to reach pH 11. Then transfer 250 ml of it to a new beaker. Add 50 mg of Amphotericin B. Mix to dissolve as much as possible. Note: Amphotericin B will not dissolve completely. Filter-sterilize the solution and store at -20oC with expiration in 183 days.
Polymyxin B stock solution. Dissolve 0.5 g of Polymyxin B in 15 ml of Distilled water completely. Filter-sterilize the solution and store at 2-8oC with expiration in 183 days.
Carbenicillin stock solution. Dissolve 0.5 g of Carbenicillin in 10 ml of Distilled water completely. Filter-sterilize the solution and store at 2-8oC with expiration in 183 days.