How to make (your own) Mueller Hinton Agar

(taken from CultureMedia Manual book)

Mueller Hinton Agar is recommended for antibiotics sensitivity test and the primary isolation of gonococci and menigococci. Also it is popularly used as a plate to cultivate Nesseria sp. Making own cultures as compared to store-bought mixes, has many advanteges, which reliability being its major advantages as commercial products might not be as standardized.


Formula, in g per L:

Steps: 

1. Add all of the above in 1L of distilled water and mix well. (pH 7.4±0.2 at 25°C)
2. Heat, while agitating frequently and let boil for about 1 minute.
3. Dispense and sterilize in autoclave at 121°C, 12-15psi for 15 mins.
4. Cool to 40-50°C and add defibrinated blood if desired.
5. If the media is used for Nesseria growth, the blood mixture should be heated to 80°C for 10 mins until chocolated, but do not overheat.

Standardization: 

This medium complies with the NCCLS (National Commitee for Clinical Lab Standard) and prepared to contain low amount of tymidine and enough calcium and magnesium ions for growth. The performance is specified by the NCCLS by conducting sensibility tests against different antibiotics using type cultures and observed after 24 hours. The diameter in mm should be as the table below:

 Well, that's all :) I'm just writing here because I can't download them due to scribd error.