摘要
[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.
出处
《安徽农业科学》
CAS
北大核心
2009年第12期5381-5382,5607,共3页
Journal of Anhui Agricultural Sciences
基金
贵州省遵义市科技局资助项目(遵市科合社字2007-21号)
关键词
细菌
DNA提取
聚合酶链反应
Bacteria
DNA extraction
Polymerase chain reaction