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本文报道了基因探针在大肠杆菌中的扩增与提纯方法。含基因探针的质粒用氯化钙方法转染大肠杆菌,被转染的细菌与质粒在培养液中同步扩增,然后用碱裂解法抽提扩增后的质粒。结果表明:(1)质粒的转化效率与其浓度有关;(2)经转染、扩增与抽提后,获得了大量的纯化质粒DNA;(3)纯化后的基因探针能满意地用于DNA 标记及分子杂交。
This article reports the amplification and purification of gene probes in E. coli. The plasmid containing the gene probe is transfected into E. coli by calcium chloride method. The transfected bacteria and the plasmid are simultaneously amplified in the culture solution, and then the amplified plasmid is extracted by alkaline lysis method. The results showed that: (1) the efficiency of plasmid transformation was related to its concentration; (2) a large amount of purified plasmid DNA was obtained after transfection, amplification and extraction; (3) the purified gene probe could be used satisfactorily DNA labeling and molecular hybridization.