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利用聚合酶链反应扩增沙门氏菌染色体DNA。

Amplification of Salmonella chromosomal DNA using the polymerase chain reaction.

作者信息

Nguyen A V, Khan M I, Lu Z

机构信息

Department of Pathobiology, College of Agriculture and Natural Resources, University of Connecticut, Storrs 06269-3089.

出版信息

Avian Dis. 1994 Jan-Mar;38(1):119-26.

PMID:8002879
Abstract

A Salmonella-specific polymerase chain reaction (PCR) was developed and standardized. The origin of the primers was a recombinant clone (C7) that contained Salmonella-specific HindIII fragment DNA of 2.1-kilobase pairs. Based on the sequence data of Salmonella enteritidis recombinant clone C7, two primers designated NK1 (21 nucleotides) and NK2 (24 nucleotides) were synthesized for use in the PCR. A Salmonella-specific 2.0-kilobase pair DNA product was amplified by the primers from 23 species of Salmonella, but not from 19 enteric and non-enteric bacteria. As little as 330 fg of Salmonella DNA was detected using either ethidium bromide/ultraviolet exposure of gels or Southern blot hybridization with a C7 clone.

摘要

开发并标准化了一种沙门氏菌特异性聚合酶链反应(PCR)。引物源自一个重组克隆(C7),该克隆包含2.1千碱基对的沙门氏菌特异性HindIII片段DNA。根据肠炎沙门氏菌重组克隆C7的序列数据,合成了两条引物,分别命名为NK1(21个核苷酸)和NK2(24个核苷酸),用于PCR。引物从23种沙门氏菌中扩增出了一条2.0千碱基对的沙门氏菌特异性DNA产物,但未从19种肠道和非肠道细菌中扩增出该产物。使用溴化乙锭/凝胶紫外线曝光或与C7克隆进行Southern印迹杂交,可检测到低至330 fg的沙门氏菌DNA。

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