双峰驼瘤胃微生物总DNA的提取与纯化

Extraction and Purification of DNA from Rumen Microbes in Bactrian Camel

【目的】提取高质量的双峰驼瘤胃微生物总DNA,为采用免培养技术研究双峰驼瘤胃微生物奠定基础。【方法】采集双峰驼瘤胃内容物,用酶解(溶菌酶、蛋白酶K)、冻融和表面活性剂十二烷基磺酸钠(sodiumdodecyl sulfate,SDS)、溴化十六烷三甲基铵(Cetyltrimethyl Ammonium Bromide,CTAB)多种处理相结合,提取微生物总DNA,并用Omega Gel Extraction kit纯化。【结果】提取到的瘤胃微生物总DNA片段大于23 kb,纯化后的DNA可用于PCR,酶切及克隆等进一步的分子生物学研究。【结论】用该提取方法得到的总DNA能够满足分子生物学试验的要求。

[ Objective] To get the high quality of total DNA of rumen microbes in Bactrian camel for molecular research with uncultured method. [ Method ] Enzyme digested （ lysozyme, proteinase K ）, frozen - thaw treatment, sodium dodecyl sulfate solution（SDS）, cetyhrimethyl ammonium bromide（CTAB） were used to treat rumen microorganisms in Bactrian camel, and purified by using Omega Gel Extraction kit . [ Result] The molecular size of the DNA extracted from the sample was much greater than 23 kb, the purified DNA can be used for PCR, and for further research of molecular biology such as cloning, enzymatic digestion. [ Conclusion] It could be concluded that the total DNA extracted by this method would satisfy the need of the research.