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常用细胞房操作
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==转染及细胞给药== #转染复合物的制备 ##由于转染试剂易被微生物及酶降解。所以转染操作需在cell房使用无菌无酶吸头进行操作。根据转染试剂的说明书,筛选DNA和转染试剂的比例 ##转染产物进行琼脂糖凝胶电泳,成功转染的产物为带正电,无琼脂糖凝胶电泳移动行为。 ##根据琼脂糖凝胶电泳结果,确认DNA和转染试剂的最佳比例。 #转染细胞 ##在前一天完成细胞铺板,第二天显微镜确认细胞密度达到70~80% ##更换培养基为新鲜培养基以及给与目标浓度的转染复合物,可预先将药物直接与培养基混匀,一次性添加(n>=2)。 #备注: ##12孔板每孔中Mate转染试剂的净含量不要超过4uL,避免细胞毒性。 ##试剂加至底部,避免附壁。 ##无特殊情况,不额外考察耗材对DNA的吸附。 ##该转染试剂使用酚红作为稀释剂,目的是便于后续直接转染cell。
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