GRADIENT MAKER, 500 ML

碱裂解法大量提取质粒（500mL菌液）

相关专题 成功走出第一步：DNA提取 碱裂解法 大量提取质粒(500mL菌液) 1、取培养至对数生长后期(一般培养12～16小时)的含待提质粒的细菌 培养液于离心杯中，配平，4℃下4000rpm离心15min，弃上清，然后将离心管倒置于干净的约纸上使上清液全部流尽。 2、向盛有细菌 沉淀物的离心管中加入20mL冰冷的溶液Ⅰ，重悬沉淀(先加5mL溶液Ⅰ，用干净的玻璃棒搅拌均匀后，再加剩下的部分)。

Sucrose Density Gradient Fractionation

and bring to 45 ml with SK (this recipe should be adjusted for the appropriate number of gradients--we want to conserve zymolyase--it's expensive)Shake gently,45 min,60 rpm,30℃FROM HERE ONOUT KEEP EVERYTHING AT 4℃Centrifuge 500 x g,10 min.4℃.Wash 1 x

Purification of Plasmid from 50 ml-culture

1. Shake E. coli harboring plasmid at 37 C overnight in 50 ml of TB containing appropriate antibiotics. (when using ampicillin, addition of the antibiotics to 100-200 ug/ml rather than usual 50 ug/ml may improve the yield of plasmids.)