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| 購買進口儀器、試劑和耗材——就在始于2001年的畢特博生物 www.hcnanjing.cn |
以下是一份GIBCO的低糖DMEM粉劑配制說明(普通高糖的DMEM培養基配法是一樣的)以及李玲、李雪峰編著的《細胞生物學實驗》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《細胞生物學實驗》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基，加入約終體積一半的三蒸水中;若配制一個包裝的培養液，在將整個包裝的干粉倒入三蒸水后，需用水洗包裝袋內面2次，倒入培養液中，以保證所有干粉都溶解成培養液。磁力攪拌或人工攪拌使之完全溶解。 3 根據包裝袋上的要求補加所需量的碳酸氫鈉;根據實驗需要，添HEPES(5-20mmol/L)、谷氨酰胺和其他特殊物質。 4 加水定容到終體積。 5 必要時用1 mol/L 鹽酸和1 mol/L 氫氧化鈉調節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。 配制好的培養液用前加入100U/mL青霉素和100U/mL鏈霉素，并根據需要加入血清(5%-20%)。 |
購買進口儀器、試劑和耗材——就在始于2001年的畢特博生物
www.hcnanjing.cn |
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