Mmushi T MSc (Microbiology).pdf

Preparation
p-iodonitrotetrazolium chloride of 0.2 g was weighed and added to a dark brown bottle
and 100 ml of distilled water to the bottle. The mixture was vigorously shaked until all
the p-iodonitrotetrazolium chloride has dissolved in the distilled water. The INT solution
was stored in the fridge until needed.
6.2. Appendix B: Media preparation
6.2.1. Preparation of 1000 ml Middlebrook 7H9 broth
4.7 g Middlebrook 7H9 Broth (Difco Cat. No. 271310
100 mL of Middlebrook ADC Enrichment (Sodium Chloride - 0.85 g, Bovine Albumin
(Fraction V) - 5.0 g, Dextrose - 2.0 g, Catalase - 3 mg)
0.5 ml Polysorbate 80 or Tween 80
Preparation
Middlebrook 7H9 Broth powder of 4.7 g was suspend in 900 mL of purified water
containing 0.5 ml polysorbate 80. Medium was sterilized by autoclaving for 15 minutes
at 121 o C and 1 atm. Aseptically 100 mL of filter sterilized Middlebrook ADC Enrichment
was added to the medium when cooled to 45°C. For preparation of agar medium 19 g of
agar bacteriological (Biolab, Batch No. 1020577) was included before sterilization.
6.2.2. Preparation of 1000 ml Luria Bertani broth
10 g Tryptone
5 g Yeast extract
5 g NaCl
Preparation
All the components were added in one liter of distilled water and left to boil to dissolve
the components completely. Medium was sterilized by autoclaving for 15 minutes at
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