农杆菌介导的转化所用基本培养基配方.doc
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农杆菌介导的转化所用基本培养基配方.doc

农杆菌介导的转化所用基本培养基配方.doc

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农杆菌介导的转化所用基本培养基配方一、NB培养基配方(基本培养基)N6大量:(mg/l)母液:10X,1L(mg)硝酸钾KNO3硫酸氨(NH4)2SO4磷酸二氢钾KH2PO4硫酸镁MgSO4·7H2O氯化钙CaCl2·2H2O2830463400185166283004630400018501660B5微量:(mg/l)母液:100X,1L(mg)硼酸H3BO4硫酸锰MnSO4·H2O硫酸锌ZnSO4·7H2O碘化钾KI钼酸钠Na2MoO4·2H2O硫酸铜CuSO4·5H2O氯化钴CoCl2·6H2O37.5820.750.250.0250.02530075820075252.52.5铁盐:(mg/l)母液:100X,1L(mg)硫酸亚铁FeSO4·7H2O乙二胺四乙酸二钠Na2EDTA27.837.327803730肌醇:(mg/l)母液:100X,1L(mg)肌醇Myo-inositol10010000有机成分:(mg/l)母液:50X,1L(mg)盐酸硫胺素ThiamineHCl盐酸吡哆醇PyridoxineHCl尼克酸Niacin水解酪蛋白Casaminoacids谷氨酰胺Glutam脯氨酸Prolineine甘氨酸Glycine101130025050025005050150001250025000100蔗糖Sucrose30000mg/l琼脂Phytagel2400mg/lpH5.8-5.9注意:有机成分不能高压灭菌,须用滤器抽滤,分装后-20℃保存二、AAM培养基配方大量:(mg/l)母液:10X,1L(mg)磷酸二氢钾KH2PO4硫酸镁MgSO4·7H2O氯化钾KCl氯化钙CaCl2·2H2O170370294044017003700294004400微量:(mg/l)母液:100X,100ml(mg)硫酸锰MnSO4·H2O钼酸钠Na2MoO4·2H2O硼酸H3BO4硫酸锌ZnSO4·7H2O碘化钾KI硫酸铜CuSO4·5H2O氯化钴CoCl2·6H2O7.580.25320.750.03870.02575825300200753.872.5铁盐:(mg/l)母液:100X,1L(mg)硫酸亚铁FeSO4·7H2O乙二胺四乙酸二钠Na2EDTA27.837.327803730肌醇:(mg/l)母液:100X,1L(mg)肌醇Myo-inositol10010000维生素:(mg/l)母液:100X,1L(mg)盐酸硫胺素ThiamineHCl盐酸吡哆醇PyridoxineHCl尼克酸Niacin0.50.50.5505050氨基酸:(mg/l)甘氨酸Glycine精氨酸Arginine谷氨酰胺Glutamine7.5174876水解酪蛋白Casaminoacids500mg/l乙酰丁香酮Acetosgringone100μΜ葡萄糖Glucose68500mg/l蔗糖Sucrose30000mg/lpH5.2注意:AAM培养基不能高压灭菌,须用滤器抽滤,分装后-20℃保存三、YEB固体培养基(pH7.0)酵母提取物蛋白胨牛肉膏蔗糖硫酸镁添加:Kan50mg/LRif50mg/L1.0g/L5.0g/L5.0g/L5.0g/L0.5g/L四、LB固体培养基(pH7.0-7.2)酵母提取物蛋白胨NaCl添加:Kan50mg/LRif50mg/L5g/L10g/L10g/L五、MS培养基配方大量:(mg/l)母液:20X,1L(mg)硝酸铵NH4NO3硝酸钾KNO3磷酸二氢钾KH2PO4硫酸镁MgSO4·7H2O氯化钙CaCl2·2H2O165019001703704403300038000340074008800微量:(mg/l)母液:100X,1L(mg)硫酸锰MnSO4·4H2O硫酸锌ZnSO4·7H2O硼酸H3BO4碘化钾KI钼酸钠Na2MoO4·2H2O硫酸铜CuSO4·5H2O氯化钴CoCl2·6H2O22.38.66.20.830.250.0250.025223086062083252.52.5铁盐:(mg/l)母液:100X,1L(mg)硫酸亚铁FeSO4·7H2O乙二胺四乙酸二钠Na2EDTA27.8537.2527853725肌醇:(mg/l)母液:100X,1L(mg)肌醇Myo-inositol10010000维生素:(mg/l)母液:100X,1L(mg)盐酸硫胺素ThiamineHCl盐酸吡哆醇P