IL-2质粒DNA探针制备的试剂与仪器
IL-2 质粒DNA探针制备的试剂与仪器 (1)含IL-2 DNA探针的质粒以及宿主菌 (2)STE 溶液:0.1mol/L NaC1 (3)Tris HC1(PH8.0):10mmo1/L、1mmo1/L (4)溶液I 50mmo1/L 葡萄糖 25mmo1/L Tris HCL(PH 8.0) 10mmo1/L EDTA(PH 8.0) 可配100ml,高压灭菌15min,贮存于4℃ (5)溶液II 0.2mo1/L NaOH(临用前用10mo1/L贮存液稀释) 1%SDS(配20%贮存液) 盖紧瓶盖,颠倒离心瓶数次,以充分混匀内容物,于室温放置5-10min. (6)溶液III 5mol/L乙酸钾 60ml 冰乙酸 11.5ml 水 28.5ml 所配成的溶液对钾是3mol/L,对乙酸根是5mo1/L。 (7)含相应抗生素的LB增养基 (8)氯霉素(0.25g/2ml)→终浓度170μg/ml (9)溶菌酶(10mg/ml,溶于10mm01/L Tris HCL PH8.0)1 ml (10)无水乙醇(部分-20℃预冷),异丙醇,75%乙醇(部分4℃预冷) (11)TE缓冲液(PH8.0) (12)5 mol/L LiC1 1.5ml (13)RNA酶 (14)3mol/L NaAC, (15)饱和酚 (16)氯仿:异戊醇(24:1) (17)质粒提取试剂盒(Promega:Wizard Minipreps DNA,NO117) (18)低温高速离心机 (19)恒温摇床
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