用溶菌酶—SDS微量法提取金黄色葡萄球菌质粒DNA

EXTRACTION OF PLASMID DNA IN STAPHYLOCOCCI BY LYSOZYME-SDS METHOD

摘要: 本文报道了检测金黄色葡萄球菌质粒DNA的微量方法(溶菌酶-SDS微量法),无需使用溶葡萄球菌素(Lyostaphin)。方法用少量的斜面培养物,经溶菌酶、SDS、EDTA裂解细胞,碱酚和氯仿-异戊醇一次脱蛋白,乙醇沉淀DNA,便得到可以直接用于琼脂糖凝胶电泳检测的质粒DNA。将该法与常规的溶葡萄球菌素法进行比较,分别检测了30株从南京地区临床分离的耐药性金黄色葡萄球菌。两种方法对细胞的裂解率为100%,质粒检出率为96.6%。用两种方法所得到的质粒电泳图谱基本相同,有些菌株发现有大质粒带。

Abstract: A simple micro-volume procedure is described for detecting the plasmid DNA in Staphylococcus aureus. The cleared lysates were prepared by iysozyme-SDS-EDTA-medieted lysis of cells obtained from a small amount of 16 hours slant culture, followed by the dep