(课件)-分子生物学实验Experimentsofmolecularbiology.pptVIP

分子生物学实验Experiments of molecular biology ;分子生物学实验室要求;实验一核酸的提取（质粒DNA的提取和纯化）;【实验原理】; 【实验材料和用品】 恒温培养箱、恒温摇床、台式离心机、高压灭菌锅、Enpendorf管、含pET-29a质粒及含目的基因质粒的大肠杆菌、碱裂解溶液，酚、氯仿、乙醇、TE缓冲液、胰RNA酶; 【实验步骤】 1. Pick single colony and inoculate 5 ml of LB broth containing 200 g/l Ampicillin or 1mg/5ml with a loosened cap and a piece of tape to hold it in place. Shake at 250 RPM 37oC overnight. 2. Centrifuge 1.5mL cells in 1.5 mL Eppendorf tube at top speed for 1 minute. Discard supernatant.? 3. Resuspend cell pellet in 100 ul of GTE buffer (50 mM Glucose, 25 mM Tris-Cl, 10 mM EDTA, pH 8).? Vortex gently if n