Alicyclobacillus Agar Recipe

1. Dissolve the following in 500ml of distilled water to make trace elements solution:
   • Na₂MoO₄ x 2 H₂O 0.03 g
   • NiCl₂ x 6 H₂O 0.02 g
   • CuCl₂ x 2 H₂O 0.01 g
   • CoCl₂ x 6 H₂O 0.20 g
   • H₃BO₃ 0.30 g
   • MnCl₂ x 4 H₂O 0.03 g
   • ZnSO₄ x 7 H₂O 0.10 g
2. Dissolve the following in 1L of distilled water to make Solution A:
   • KH₂PO₄ 3.0 g
   • Glucose 5.0 g
   • Yeast extract 2.0 g
   • (NH₄)₂SO₄ 0.2 g
   • MgSO₄ x 7 H₂O 0.5 g
   • CaCl₂ x 2 H₂O 0.25 g
3. Dissolve the 12g of agar in 500ml to make Solution B.
4. Autoclave solutions separately.
5. To make final agar combine solution A with solution B and aseptically add 1ml of trace elements solution.