使用说明：
1. 对于贴壁细胞：
   a) 将细胞培养于培养皿或培养板内，通常在铺板后1-2天内长到70-80％满为宜。后续说明都按6孔板内
      一个孔进行说明，如果有更多个孔或大小不相同的培养器皿，请自行换算。
   b) 在转染前30-60分钟，吸去细胞培养液，加入新鲜的不含抗生素的完全培养液2毫升。注意：转染时
      最好使用新鲜配制且pH值经过精心调校的培养液；转染用的培养液可以在配制后分装冻存，使用时再
      解冻。
   c) 取2-6微克待转染的质粒DNA (质粒总体积不宜超过20微升)，加入到100微升氯化钙溶液中，混匀。
   d) 把DNA-氯化钙溶液加入到100微升BBS溶液中，混匀，室温孵育10-20分钟。此时不会产生可见的沉
      淀。
   e) 把DNA-氯化钙-BBS混合物均匀滴加到整个6孔板内。在含5%二氧化碳的37℃细胞培养箱内培养。
   f) 根据实验要求和磷酸钙对于不同细胞的毒性不同，在4-16小时后轻轻晃动培养板数次以充分悬浮一些
      磷酸钙沉淀，吸去含磷酸钙沉淀的培养液，加入2毫升新鲜的完全培养液，继续培养。
   g) 通常在转染约24小时后就可以检测到转染基因的表达。
2. 对于悬浮细胞：
   a) 离心收集悬浮细胞，用PBS洗涤一次。
   b) 按照上面的步骤c)和d)制备DNA-氯化钙-BBS混合物。
   c) 每106个细胞的沉淀，用100微升DNA-氯化钙-BBS混合物重新悬浮，室温放置20分钟。
   d) 在一个6孔板孔内加入2毫升完全培养液，然后加入来自上一步的细胞-DNA-氯化钙-BBS混合物，混
      匀。
   e) 在含5%二氧化碳的37℃细胞培养箱内培养。
   f) 根据实验要求和磷酸钙对于不同细胞的毒性不同，在4-16小时后离心收集细胞，用PBS洗一次，然后
      用2毫升完全培养液重新悬浮细胞，继续培养。
   g) 通常在转染约24小时后就可以检测到转染基因的表达。

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