Protocol for Middlebrook 7H11 Agar (Difco) with Glycerol
Material: Middlebrook 7H11 Agar Powder, Glycerol, 20% Tween 80, OADC
1. Take 900 ml distilled water in a glass flask
2. Place the flask on a magnetic stirrer
3. Add 21 g of the powder in water and stir it
4. Add 5 ml of glycerol (Pipette slowly) and stir it
5. Autoclave the medium at 121 0C for 15 minutes
6. After Autoclaving cool the medium up to 50 0C (The temperature of container which can be tolerated by skin).
7. Then, aseptically add 100 ml of OADC to the medium*
8. Add 2.5 ml of 20% Tween 80 to the medium *
Material: Middlebrook 7H11 Agar Powder, Glycerol, 20% Tween 80, OADC
1. Take 900 ml distilled water in a glass flask
2. Place the flask on a magnetic stirrer
3. Add 21 g of the powder in water and stir it
4. Add 5 ml of glycerol (Pipette slowly) and stir it
5. Autoclave the medium at 121 0C for 15 minutes
6. After Autoclaving cool the medium up to 50 0C (The temperature of container which can be tolerated by skin).
7. Then, aseptically add 100 ml of OADC to the medium*
8. Add 2.5 ml of 20% Tween 80 to the medium *
* Steps 7 and 8 must be done in a safety hood
* Temperature higher than 500C can denature OADC and Tween 80, on the other hand low temperature will cause solidification of agar. Therefore, add OADC and Tween 80 at 500C and pour the medium on plates as soon as possible before solidification.
* Temperature higher than 500C can denature OADC and Tween 80, on the other hand low temperature will cause solidification of agar. Therefore, add OADC and Tween 80 at 500C and pour the medium on plates as soon as possible before solidification.
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