热变性温度法测定细菌DNA中G+C的含量

DETERMINATION OF THE GUANIME PLUS CYTOSINE CONTENT OF BACTERIAL DNA FROM ITS THERMAL DENATURATION TEMPERATURE

近年来,随着分子生物学的不断发展,为细菌分类鉴定开辟了许多新途径,提供了许多新技术,如细菌脱氧核糖核酸(DNA)碱基成份分析、核酸分子杂交等。测定细菌DNA中鸟嘌呤(G)和胞嘧啶(C)的克分子百分数(Mol%)的方法,在细菌分类鉴定中是一种新方法。

This paper reports our work of the use of the thermal denaturation temperature(T)method to determine G+C Mol%of bacterial DNA.The method is simple,convenient and apparently more precise.Experimental details of DNA extraction and purification,Tm determination and calculation of G+G Mol%have been described.We have improved the semiconductor thermistor thermometer.The temperature of the DNA solution during an experiment was measured directly in the cell with the probe of a thermistor thermometer inserted through the stopper.This may insure the accuracy and reproducibility of experimental results.When 77 strains of 13 bacterial species were determined as G+C Mol%,it was shown that their G+C content was approximately equal to those of reported in literature.This method is now being used to identify ambiguous strains in our laboratory.