Peeling Away Problems

The Antioxidating, Antimicrobial, and Antimutagenic Effects of Tree Bark

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Ames Test

(See Reference 4 for details)

1.  Make minimal agar plates and top agar.

2. Grow bacteria culture (Salmonella typhimurium TA 1535) overnight.  

3.  Make a mixture of 100 uL bacterial solution, a variable(s) (control, or NaN3, or extract, or NaN3+extract), and 3 mL of top agar.

4. Pour mixture onto minimal agar plate, swirl, and let incubate for 2 days.

 

Top Agar 

       0.6  g       Bacto agar 

       0.5  g       Sodium Chloride (NaCl) 

       100 mL    ddH2O

       Melt in steam kettle and sterilize 

       Add 10mL Biotin/Histidine Solution 

Biotin/Histidine Solution 

        30.9mg       D-Biotin (C10H16N2O3S)

       24   mg       L-Histidine (C6H9N3O2)

 

Minimal Agar Plates 

       7.5  g       Bacto agar 

       455 mL    ddH2O

       autoclave  to sterilize, then add sterilized 

       20   mL       25X Vogel-Bonner Salts 

       25   mL       40% glucose 

Vogel-Bonner Salts (25X) 

        670  mL  warm ddH2O

        5     g       MgSO3·7H2O

        50   g       monohydrate citric acid (C6H8O7)

        250 g       anhydrous potassium phosphate dibasic (K2HPO4)  

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