Heart Infusion Broth with Porcine Serum and Fresh Yeast Extract

[所属分类:培养基配方] [发布时间:2021-4-1] [发布人: ] [阅读次数:] [返回]

Heart Infusion Broth with Porcine

Serum and Fresh Yeast Extract

培养基配方

Composition per liter:
Beef heart, infusion from..........................................................500.0g
Tryptose......................................................................................10.0g
NaCl..............................................................................................5.0g
Porcine serum, inactivated.....................................................200.0mL
Fresh yeast extract (25% w/v solution) .................................100.0mL
pH 7.5 ± 0.2 at 25°C
Porcine Serum:
Composition per 200.0mL:
Porcine serum ........................................................................200.0mL
Preparation of Porcine Serum: Adjust the pH of 200.0mL of porcine serum to 4.4 with sterile 1N HCl. Do not overshoot pH below 4.2. Allow serum to stand at 4°C for 18–20 hr. Adjust pH to 7.0 with sterile NaOH. Aseptically centrifuge at 9000 rpm for 20 min. Discard sediment. Filter supernatant solution through a 0.85μm filter. Filter sterilize through a 0.22μm filter. Store at −70°C.
Fresh Yeast Extract Solution:
Composition per 100.0mL:
Baker’s yeast, live, pressed, starch-free......................................25.0g
Preparation of Fresh Yeast Extract Solution : Add the live Baker’s yeast to 100.0mL of distilled/deionized water. Autoclave for 90 min at 15 psi pressure–121°C. Allow to stand. Remove supernatant solution. Adjust pH to 6.6–6.8.
Preparation of Medium: Add components, except porcine serum and fresh yeast extract solution, to distilled/deionized water and bring volume to 700.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C. Aseptically add sterile porcine serum and fresh yeast extract solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Acholeplasma species.