菌株简介: TG1来源于E. coli K-12菌株,是目前生长速度最快的克隆用大肠杆菌菌株,在平板上37℃,7h可见克隆。主要的噬菌体展示用菌株,同时也可用于普通质粒的构建,lacIqZΔM15的存在使其可以用于蓝白斑筛选等实验;但不含核酸酶endA1突变,体内核酸酶含量较高,提取质粒时推荐使用质粒提取试剂盒中去蛋白液以去除菌体内大量的核酸酶。TG1菌株需在无抗生素的LB中,在37℃有氧的条件下培养,然后使用20%的甘油保藏菌种。 4. Minimal Medium plate配制方法:Prepare stocks of the following 1 M MgCl2•6H2O: Dissolve 20.33 g in distilled water to a final volume of 100 ml and autoclave. 1 M CaCl2•2H2O: Dissolve 14.7 g in distilled water to a final volume of 100 ml and autoclave. 1 M thiamine hydrochloride: Dissolve 33.73 g in distilled water to a final volume of 100 ml and sterilize using a 0.22 μm filter. 20% glucose: Dissolve 20 g of D-(+)-glucose (anhydrous) in distilled water to a final volume of 100 ml and sterilize by filtration through a 0.2 μm filter. Do not autoclave. In a 500 ml bottle, dissolve 6 g of Na2HPO4 (dibasic), 3 g of KH2PO4(monobasic) and 1 g of NH4Cl in distilled water to a final volume of 500 ml. adjust the pH to 7.4 with NaOH. In a separate 1 liter bottle, add distilled water to 15 g of Bacto-agar to a final Volume of 500 ml. Autoclave both bottles simultaneously to sterilize. Cool both bottles to 50-60°C and combine. Add1 ml of 1 M MgCl2•6H2O, 1 ml of 1 M CaCl2•2H2O, 1 ml of 1 M thiamine hydrochloride and 5 ml of 20% glucose. Pour plates immediately.