提取鼠肠道内微生物基因组DNA的方法研究

通过改进鼠肠道微生物基因组DNA提取方法以期更全面地反映鼠肠道茵群的真实情况.采用CTAB和SDS结合的方法裂解细胞,同时改进了传统的酚/氯仿抽提方法,提取全过程控制在2 h以内.通过紫外分光光度计,细菌通用引物PCR,总菌群实时定量PCR,变性梯度凝胶电泳(DGGE)对改进方法及两种试剂盒法提取DNA的结果进行比较,评价了所建立的高效提取方法.与试剂盒相比,改进的方法DNA得率较高,简便快速,成本低廉.同时后续的PCR鉴定及DGGE菌群多样性分析显示,改进的方法可以更好地揭示鼠肠道菌群分布和特性.

An Effective Method for Genomic DNA Extraction from Rat Intestinal Microflora

A modified method for extracting microbial community DNA from rat intestinal samples to fully reflect the real situation of rat intestinal microflora is described in this paper. Combination of CTAB and SDS was used for cell lysis, the traditional DNA extraction method was improved, and the whole process was controlled within two hours. This modified method and other two DNA extraction kits were evaluated by UV spectrophotometer, bacterial u-niversal primer PCR, real -time quantitative PCR and denaturing gradient gel electrophoresis (DGGE). Results show the modified method has a higher yield of DNA and it is simple, rapid and cost - effective compared with the other two kits. The PCR and DGGE show that the modified method can reveal the distribution and trait of rat intesti-nal microflora better.