Mueller Hinton II (Cation Adjusted)
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Purpose
Mueller Hinton II broth is used for quantitative susceptibility testing for gram-negative and gram-positive facultative and aerobic bacteria. The cation adjustment is necessary for the in vitro testing of Pseudomonas aeruginosa.
Materials
- 3.0g/L Beef Extract
- 17.5g/L Acid Hydrolysate of Casein
- 1.5g/L Starch
- 20-25mg/L Ca2+
- 10-12.5mg/L Mg2+
- dH2O
Procedure
- Create a stock solution of Ca2+ by adding 8.36g of MgCl2·6H2O into 100mL dH2O and sterile filter solution resulting a final concentration of 10mg/mL.
- Create a stock solution of Mg2+ by adding 3.68g of CaCl2·2H2O into 100mL dH2O and sterile filter solution resulting a final concentration of 10mg/mL.
- In a 1L bottle add 3.0g Beef Extract, 17.5g Acid Hydrolysate of Casein, .5g Starch to 997mL of dH2O
- Autoclave media at 121°C for one hour
- After autoclave cycle is complete, allow media to cool down to 50°C before adding 1mL of Mg2+ stock and 2mL Ca2+
- for procedure see MIC P. aeruginosa