1. Measrue out 5% less distilled water than desired total volume of medium using a
mixing container that is as close to the final volume as possible.
2. Add powdered medium to 15 to 30℃ (room temperature)water with gentle stirring.
(Do not heat water)
3. Rinse out inside of package to remove all traces of powder.
4. Add 3.7g of NaHCO3 per liter of medium.
5. Dilute to a desired volume with water. Stir until dissolved. (Do not over-mix)
6. Adjust pH of medium to 0.2-0.3 below desired final working pH* use of IN NaOH or
IN HCl is recommended. (Add slowly with stirring) After pH has been adjusted keep
container closed until medium is filtered.
7. Sterilize immediately by membrane filtration. (Positive pressure recommended)
*pH unite will usually rise 0.1-0.3upon filtration.
1 制备新鲜三蒸水或Millipore超纯水。
2 称取所需量的干粉培养基，加入约终体积一半的三蒸水中;若配制一个包装的培养液，在将整个包装的干粉倒入三●蒸水后，需用水洗包装袋内面2次，倒入▂培养液中，以保证所有干粉都溶解成培养液。磁力搅拌或人工搅拌使之完全溶解。
3 根据★包装袋上的要求补加所需量的碳酸氢钠;根据实验需要，添HEPES(5-20mmol/L)、谷氨酰胺和其他特＠殊物质。
4 加水定容到终体积。
5 必要时用1 mol/L 盐酸和1 mol/L 氢氧化钠调节pH。
6 用无菌0.22um滤膜过滤除菌，分装于无菌血清瓶中，4℃冰箱保存。
配制好的培♀养液用前加入100U/mL青霉素和100U/mL链霉素，并根据需要加入血清(5%-20%)。
