使用说明：
1.样品收集
  a)对于组织样品：
    切下组织，并剪切成小块，置液氮中冻结，研碎或捣碎。或直接冰浴上匀浆。
  b)对于贴壁细胞：
    胰酶消化后，PBS或生理盐水洗一次，1000-2000g离心1-2分钟，弃上清，收集细胞。
  c)对于悬浮细胞：
    1000-2000g离心1-2分钟，弃上清，收集细胞。
2.DNA ladder抽提
  a)每1毫升样品裂解液中加入5微升蛋白酶K，混匀。
  b)对于上述收集好的样品，每5毫克组织或者10⁶个细胞中加入500微升添加了蛋白酶K的样品裂解液，
    Vortex混匀，充分裂解组织或细胞。
  c)50℃水浴消化过夜(通常12-20小时皆可)。
  d)加入500微升Tris平衡苯酚。
  e)Vortex剧烈混匀，使有机相和水相充分混合，以达到抽提效果。4℃，12,000g离心5分钟。
  f)缓慢吸出酚相及中间相(可以吸除少量靠近中间相的水相液体)，剩余的水相用等体积Tris平衡酚再抽提
    一次(同步骤e)。
  g)缓慢吸出酚相及中间相(可以吸除少量靠近中间相的水相液体)，剩余的水相用等体积氯仿再抽提一次
    (同步骤e)。
  h)慢慢吸出约300微升上清液，加入60微升10M醋酸铵和600微升无水乙醇，颠倒数次混匀，此时可见DNA沉
    淀产生。-20℃冻存1小时，以充分沉淀小片断DNA。冻存过夜或-70℃冻存效果更佳。
  i)4℃，12, 000g 离心10分钟，弃上清。
  j)加入600微升70%乙醇，轻轻颠倒约2次。4℃，12, 000g 离心10分钟，小心吸去上清。注意：70%乙醇
    洗涤的时候，千万注意避免损失一些细小的DNA沉淀，这些沉淀中大部分是你所需的DNA ladder。
  k)尽量吸除残余的乙醇，待看不到明显的液体时，立即加入50-100微升TE溶解DNA。注意：不可过分干燥
    基因组DNA沉淀，否则会极难溶解。如果发现DNA沉淀难以溶解，可以在4℃用摇床缓慢摇动过夜，以溶
    解DNA沉淀。
  l)取部分抽提得到的DNA，1%琼脂糖凝胶电泳分析。如果细胞发生凋亡，就可以观察到典型的DNA
    ladder。电泳时一定要注意换用新鲜配制的电泳液，DNA凝胶也要用新鲜配制的电泳液配制并新鲜配制
    后使用。电泳时为获取最佳的电泳效果使ladder充分分开，电泳速度宜适当慢一些，凝胶宜适当长一
    些，而加样孔宜更加扁平一些。选取适当较薄的梳齿，往往会获得更好的ladder电泳效果。

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