自行配制DMEM的方法

相关专题

细胞培养基的配制 细胞培养专题 MEM细胞培养基

以下是一份GIBCO的低糖Dmem 粉剂配制说明(普通高糖的DMEM培养基配法是一样的)以及李玲、李雪峰编著的《细胞生物学实验》中配制方法。

TO PREPARE undefinedLIQ UID

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 gen tle 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 pundefined 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)

~undefinedpH unite will usually rise 0.1-0.3upon filtration.

另外，在李玲、李雪峰编著的《细胞生物学实验》中配制方法如下：

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%)。