自我总结 质粒抽提
质粒抽提将过夜培养的菌液转入1.5ml的eppendorf管中,2,500g离心5min,弃上清,用滤纸吸尽管壁上残余的上清,加入200l resuspension solution悬浮菌体,再加入200l lysis solution混匀,之后加入200l neutralization solution混匀,12,500g离心10min,将上清转移到新的无菌eppendrof管中,加入等体积预冷的异丙醇,于-20放置10min,12,500g离心1015min,弃上清,干燥后加入100150l的双蒸水溶解质粒DNA(根据实验需要,双蒸水的量可做适度调整)。溶液配制: 细胞悬浮液(cell resuspension solution)Tris HCl PH7.5 50mMEDTA PH8.0 10mMRNAaseA 100mg/ml 细胞裂解液(cell lysis solution)NaOH 0.2MSDS 1% 中和液(Neutratization)Potassium acetate (KAC) PH 4.8 1.32M