以下是一份GIBCO的低糖DMEM粉劑配制說明(普通高糖的DMEM培養(yǎng)基配法是一樣的)以及李玲、李雪峰編著的《細(xì)胞生物學(xué)實(shí)驗(yàn)》中配制方法。

TO PREPARE 1*LIQUID

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.

另外，在李玲、李雪峰編著的《細(xì)胞生物學(xué)實(shí)驗(yàn)》中配制方法如下：

1 制備新鮮三蒸水或Millipore超純水。

2 稱取所需量的干粉培養(yǎng)基，加入約終體積一半的三蒸水中;若配制一個(gè)包裝的培養(yǎng)液，在將整個(gè)包裝的干粉倒入三蒸水后，需用水洗包裝袋內(nèi)面2次，倒入培養(yǎng)液中，以保證所有干粉都溶解成培養(yǎng)液。磁力攪拌或人工攪拌使之完全溶解。

3 根據(jù)包裝袋上的要求補(bǔ)加所需量的碳酸氫鈉;根據(jù)實(shí)驗(yàn)需要，添HEPES(5-20mmol/L)、谷氨酰胺和其他特殊物質(zhì)。

4 加水定容到終體積。

5 必要時(shí)用1 mol/L 鹽酸和1 mol/L 氫氧化鈉調(diào)節(jié)pH。

6 用無菌0.22um濾膜過濾除菌，分裝于無菌血清瓶中，4℃冰箱保存。

配制好的培養(yǎng)液用前加入100U/mL青霉素和100U/mL鏈霉素，并根據(jù)需要加入血清(5%-20%)。

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