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

  1. 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.
  2. 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.
  3. In a 1L bottle add 3.0g Beef Extract, 17.5g Acid Hydrolysate of Casein, .5g Starch to 997mL of dH2O
  4. Autoclave media at 121°C for one hour
  5. After autoclave cycle is complete, allow media to cool down to 50°C before adding 1mL of Mg2+ stock and 2mL Ca2+
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