ATCC Medium: 1561 Middlebrook w/STREP 100
Note: Middlebrook 7H10 contains agar, Middlebrook 7H9 is for broth
For Solid Media Middlebrook
7H10 (BD 262710)…………19.0 g
Glycerol……………………………………. 5.0 ml
DI Water…………………………………..850.0 ml
Add 19 grams of dehydrated Middlebrook 7H10 medium to 900 ml of water containing 5.0 ml of glycerol. Bring to a boil to dissolve and autoclave at 121 C for 30 minutes. Cool to 50-55 C.
Aseptically add 100 ml of AODC Enrichment and 100 mcg of strep 100.
Dispense as required. OADC Enrichment (see below)……100.0 ml
STREP (100 mcg/ml)………………..50.0 ml
OADC Formulation:
Add Per Liter of Middlebrook
Oleic Acid (Sigma Cat # O-1008)………………… 0.05g
Albumin Fraction V Bovine (Sigma A9647-1KG)…5.0 g
Dextrose (BD 215530)……………………………….2.0 g
Catalase, beef (Sigma C-9322)………………….0.004 g
NaCl…………………………………………………..0.85 g
Distilled water……………………………………….100 ml
For Broth
Middlebrook 7H9 Broth (271310) ………………..4.7 g
Glycerol……………………………………………...2.0 ml
Distilled water ……………………………………850.0 ml
Dissolve 4.7 grams of dehydrated Middlebrook 7H9 Broth into 900 ml of deionized water. Add 2.0 ml of Glycerol. Mix well. Autoclave at 121?C
Aseptically add 100ml of Middlebrook ADC Enrichment (see Below) Dispense as required.
ADC ……………………….100.0 ml
Strep 100 (100mcg/ml)…….50.0 ml
ADC Enrichment formula:
Bovine Serum Albumin, Frac V…………….5.0 g Dextrose……………………………………….2.0 g
Catalase (beef)……………………………….3.0 mg
DI water…………………………………….100.0 ml
Filter Sterilize.