摘要
本文参考分子生物学有关资料对我国南方家鼠寄生蚤,缓慢细蚤(Leptopsylla segnis)和不等单蚤(Monopsyllus anisus)基因组DNA的提取方法进行探索。其主要过程可分四步:(1)用玻璃匀浆器匀浆。(2)以SDS破坏细胞,其终浓度为0.8％～1％(W/V);蛋白酶K消化蛋白质,终浓度为300μg/ml。(3)苯酚、氯仿、异戊醇反复抽提。(4)得到的DNA用紫外分光光度仪和琼脂糖电泳检测。用本法所获得蚤类DNA纯度和含量基本可达到进一步实验的要求,为今后的酶切实验、探针制备、基因克隆和分子杂交等提供较理想的方法。
A method for extraction of the genomic DNA of household rat fleas, Leptopsylla segnis and Monopsyllus anisus in southeast China, has been established based on general molecular biological methods. In comparison with the general tissue and cell genomic DNA techniques, the method was different in that: (1) the working concentration of proteinase K was about 300 ug/ ml and SDS about 0.8%-1% (w / v); (2) the flea tissues must be homogenized in a glass homogenizer; (3) extraction of flea genomic DNA generally use the mix of phenol-chloroform- isoamyl; (4) flea DNA was determined by HITACHI 557 Double Wavelength Double Beam Spectrophotometer and electrophoresis.
出处
《地方病通报》
1993年第1期56-60,共5页
Endemic Diseases Bulletin
基金
福建省自然科学基金资助项目~~
关键词
蚤
DNA
分子生物学
提取
Fleas
DNA
Molecular biology
Extraction method
