常用培养基配制.docx

常用培养基LB培养基/升终1 X浓度胰蛋白胨10?g1.0% (w/v)酵母抽提物5?g0.5% (w/v)NaCl10?g1.0% (w/v)H2O至1升?调节pH值至7.0低盐LB培养基/升终1 X浓度 胰蛋白胨10?g1.0% (w/v)酵母抽提物5?g0.5% (w/v)NaCl10?g0.5% (w/v)H2O至1升?调节pH值至7.0Terrific Broth 培养基/升终1 X浓度胰蛋白胨12?g1.2% (w/v)酵母抽提物24?g2.4% (w/v)甘油4?g0.4% (w/v)加H2O至900 ml?灭菌，冷却至60°C后加入100 ml过滤灭菌的10X TB磷酸盐 (0.17 M KH2PO4, 0.72 M K2HPO4)SOB 培养基/升终1 X浓度胰蛋白胨20?g2.0% (w/v)酵母抽提物5?g0.5% (w/v)NaCl0.5?g0.05% (w/v)250?mM?KCl10?ml2.5?mMH2O至900 ml?调节pH值至7.0，加H2O至990 ml灭菌、冷却至室温。使用前加入10 ml灭菌的1 M MgCl2 溶液10?mMSOC 培养基SOB 培养基(1升)加入20 ml 过滤灭菌的1 M葡萄糖M9 基本培养基/升终1 X浓度5X M9 salts200?ml?灭菌H2O至1升?1?M?MgSO41?ml1?mM20% 葡萄糖10?ml0.2 (w/v)1?M CaCl20.1?ml0.1?mM5X M9 Salts/升终1 X浓度Na2HPO4x7H2O64?g47.8?mMKH2PO415?g22?mMNaCl2.5?g8.6?mMNH45?g18.7?mM添加剂/升终1 X浓度抗生素(如果需要)氨苄青霉素?50?μg/ml氯霉素?20?μg/ml卡那霉素?30?μg/ml四环素?12?μg/ml半乳糖苷(如果需要)X-Gal?20?μg/mlIPTG?0.1?mM培养基含有琼脂或琼脂糖琼脂(平板)15?g1.5%?(w/v)琼脂(软性琼脂)7?g0.7%?(w/v)琼脂糖(平板)15?g1.5%?(w/v)琼脂糖(软性琼脂糖)7?g0.7%?(w/v)参考文献 Atlas, R.M., Handbook of Microbiological Media, second edition, Ed. Parks, L.C., CRC Press, N.Y., 1997. Sambrook, J., et al., Molecular Cloning: A Laboratory Manual, second edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, A1.2-A2.12, 2001. 常用储存液10 M 醋酸铵醋酸铵385.4?gH2O至500?ml1 M CaCl2CaCl2x6H2O219.1?gH2O至1升100X Denhardt 溶液终浓度Ficoll 40010?g0.02%?(w/v)聚乙烯吡咯烷酮10?g0.02%?(w/v)牛血清白蛋白10?g0.02%?(w/v)H2O至500 ml?过滤灭菌后分装(25 ml)，-20°C 保存0.5 M EDTA (乙二胺四乙酸) (pH 8.0)Na2EDTAx2H2O186.1 gH2O至700 ml10 M NaOH (~50 ml)调节pH值至8.0H2O至1升10?mg/ml 溴化乙锭溴化乙锭0.2?gH2O至 20 ml充分混匀后4°C黑暗保存警告: 溴化乙锭是一种突变剂。使用溴化乙锭溶液时需带手套；称量其粉末时需带面具。1 M KClKCl74.6?gH2O至1升1 M MgCl2MgCl2x6H2O20.3?gH2O至100 ml1 M MgSO4MgSO4x7H2O24.6?gH2O至100 ml5 M NaClNaCl292.2?gH2O至1升10 M NaOHNaOH400.0?gH2O至1升1 M Tris-HCl[三羟甲基氨基甲烷盐酸盐]Tri base121.1?gH2O至800 ml用于浓缩的HCl调节至所需的pH值混合后加H2O至1升3 M 醋酸钠 (pH 5.2 或 7.0) (2)醋酸钠 x 3H2O401.1?gH2O至800 ml用醋酸调节pH值至7.0或用冰醋酸调节pH值5.2H2O至1升参考文献 Atlas, R.M., Handbook of Microbiological Media, second edition, Ed. Parks, L.C., CRC Press, N.Y., 1997. Sambrook, J., et al., Molecular Cloning: A L