赛默飞世尔Thermo Fisher HYDROXYLAMI

碱裂解法大量提取质粒（500mL菌液）

和14步对提高提取的质量和产率比较重要。 14、加入等体积的PEG-NaCl溶液，轻轻混匀，室温静置30min(大于10min)。 15、配平，4℃下8500rpm离心10min，沉淀用数mL70%乙醇洗。 16、待乙醇挥发干净后，将沉淀溶于1-5mLTE或DDW(双蒸水)中。 溶液配制 一、溶液Ⅰ配制 溶液Ⅰ组成：葡萄糖，EDTA，Tris-HCl。三种物质在溶液Ⅰ中的终浓度分别为：0.05mol/L葡萄糖，0.025mol

Purification of Plasmid from 50 ml-culture

of cold 70% ethanol. Discard the fluid. 14. Add to the pellet 400 ul of TE containing DNase-free RNase A (20 ug/ml). Incubate the tube for 30 min at 37 C. 15. After 30 min, carefully check the content of the tube. If nucleic acid pellet

BAC DNA Isolation from 200 ml Cultures by a Cleared Lysate Method

antibiotic. After incubating at 37 degrees C for 8-10 hours with 250 rpm shaking, the culture is transferred to a 500 ml flask containing 200 ml of the same medium and incubated for 8-10 hours under the same conditions. After harvesting the cells