2种改进的未知细菌DNA提取方法

Two Improved Extraction Methods of DNA in Unknown Bacteria

[目的]研究细菌DNA的提取方法，为未知细菌的研究提供方法支持。[方法]从DNA的纯度、基因组DNA的酶切效果、16SrRNA基因扩增等方面比较了细菌DNA的2种提取方法。[结果]方法Ⅰ提取的DNA的纯度（OD260／OD280）为1．51-1．65，6个样品的平均值为1．56。方法Ⅱ提取的DNA的纯度（0D260／OD280）为1．25～1．41，6个样品的平均值为1．33。方法隈取的DNA纯度较高，但其试验步骤稍微繁琐。方法Ⅱ的试验步骤稍微简单，但试验时间较长。2种方法均提取到长度大于21kb）的细菌基因组DNA。对2种方法提取的DNA进行PCR扩增都得到长度约600bp的条带，效果较好。[结论]2种方法提取的DNA质量均较高，都可以直接用于RFLP、PCR等研究。

[Objective] The aim was to study the extraction method of DNA in bacteria so as to supply methodological support for studying unknown bacteria. [Method] The 2 extraction methods of DNA in bacteria were compared in aspects such as purity of DNA, enzyme digestion effect of genome DNA and gene amplification of 16S rBNA. [Result] The purity of DNA （ OD260/0D280）extracted by method 1 was between 1.51 and 1.65 and the average purity of the 6 samples was 1.56. The purity of DNA extracted by method 2 was between 1.25 and 1.41 and the average purity of the 6 samples was 1.33. The purity of DNA extracted by method 1 was higher, but its expel/mental procedure was a little tedious. The expeimiental procedure of method 2 was a little simple, but its experimental duration was longer. The genome DNA longer than 21 kb in bateria was extracted by the 2 methods. The bands with length being about 600 by were obtained from the PCR amplification of DNA extracted by the 2 methods and their amplification effects were better. [Conclusion] The quality of DNA extracted by the 2 methods was relatively high and both of them could be used in RFLP and PCR study directly.