Study On DNA Extraction And Identification Of Intestinal Flora

With the development and exploration of microbial technology and the rise of fecal bacteria transplants in recent years,the investigation of intestinal flora are paid more attention.The intestinal flora plays a role in metabolic activities in ones body,as well as its composition and function,which has become the concerning focus of scholars at home and abroad.The effects of fecal bacteria transplants being used in clinical treatment of refractory bowel disease has been certified;and it is proved that the intestinal floras are closed linked with human diseases.Therefore,adopting which method of extraction for complete intestinal bacteria,is a prerequisite for correctly understanding intestinal flora.At present,the general method for extracting the intestinal flora from the fecal samples,however,the component of stool samples is very complicated.They contain the intestinal flora,but also includes various types of organic protein,various inorganic salts,all kinds of fiber,undigested food residues and the lining of the gut mucosal cells.Thereby,when extracting the DNA of intestinal bacteria or intestinal flora,fecal samples require some processing operations which can ensure the extractive concentration by a pretreatment while increase the purity.It takes a long time to detect the DNA of intestinal flora by sequencing and DGGE electrophoresis and the operation is very complicated.Spectroscopy as a purely physical optical method,detect fast and easy to operate,can achieve the purpose of DNA testing and identification of intestinal flora.This paper conducted a comprehensive analysis and research in influence factors of DNA purity and concentration of intestinal flora DNA extraction which include:different proportions of samples and physiological saline,centrifugal speed,centrifugal time,centrifugal times.The result shows the changing trends of DNA purity influenced by different factors.And the influence sequence of the above factors can be drawn out.So that we can choose an optimal method for preprocessing of intestinal flora DNA extraction,To the different kinds intestinal flora DNA,one spectrum detection can get all band UV absorbance information.Using principal component score values obtained by principal component analysis method,combinedwith Euclidean distance to classify different species of intestinal flora of DNA,such as dogs,cats,rabbits,has verified the feasibility of spectrometry for different species classified DNA.Establishing an optimized gut bacteria DNA extraction pretreatment method by orthogonal analysis method can get DNA of high purity and high concentration.Using the method of principal components and cluster analysis to identify different species of intestinal flora of DNA.The work in this paper provides a reference for intestinal flora or intestinal flora DNA extraction and identification.