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文献和实验稳定转染细胞所需用量需要通过实验优化。最适用量通过建立细胞死亡曲线获得。将细胞稀释至1000cell/ml,每孔100ul加入有培养基的24孔板,将每孔中的G418浓度稀释至0,100,200,300,400,500,600,700,800,900,1000,1100ug/ml等12个级别, 培养10-14天,以最低细胞全部死亡浓度为基准,一般400-800左右,筛选时比该浓度再高一个级别,维持时使用筛选浓度的一半即可。 G418选择浓度范围 细胞系或机体 G418浓度
ES Cell Culture and Manipulation
poked in it). Place the cells inside the cell-culture cooler, and place at -70°C overnight. The next day, transfer the cells to liquid nitrogen. Transformation of ES cells Preparation of targeting DNA Digest 100µg of targeting construct
1.Basic Techniques - The “Do’s and Don'ts” of Cell CultureGiven below are a few of the essential "do’s and don’ts" of cell culture. Some of these are mandatory e.g. use of personal protective equipment (PPE). Many of them are common sense and apply
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