A metagenome DNA extracting method of intestinal flora in mice for molecular diversity analysis based on PCR technology

Objective To establish a method of metagenome DNA extraction of intestinal flora in mice for molecular diversity analysis based on PCR.Method The faecal samples were collected from intestinal tracts of mice.These samples were washed by acetone,and then centrifuged in different speeds.The cells were lysed by lysozyme and SDS,treated by CTAB,and then extracted with phenol/chloroform.The metagenome DNA of bacterial species was determined by a UV spectrophotometer,agarose gel electrophoresis.The 16S rDNA was amplified from these DNA samples through a set of bacterial universal primers.The PCR products were analyzed by Amplifed Ribosomal DNA Restriction Analysis(ARDRA).Result The metagenome DNA of intestinal flora in mice extracted by this method was about 23 kb;the A260/A280 of DNA was between 1.8 and 2.0;the PCR products were about 1.4 kb and can be applied to ARDRA for further analysis.Conclusion This method is cost-effective and easy,which is suitable for metagenome DNA extraction of intestinal flora.