Human feces DNA extraction method

Abstract

The invention discloses a human feces DNA extraction method. The prior art is complicated and time-consuming; and the trace DNAs extracted by the previous sampling method is too little to perform molecular diagnosis. The extraction method uses selective sample collection PSC equipment for feces samples and adopts a low temperature preservation method. The extraction method comprises the following steps: picking a sample by using a sterilized gun head or sterilized blade and directly placing the picked sample on ice; and adding a feces lysis solution, fully cracking and evenly mixing, adding bovine serum albumin, centrifuging to remove the supernatant, adding protease K to perform a digestion reaction at 58 DEG C, then adding a protein precipitation solution to precipitate the protein, centrifuging to obtain the supernatant, adding a silica gel membrane binding solution to fully bind DNAs, centrifuging to obtain precipitates, adding a silica gel membrane washing solution to wash twice, centrifuging, and adding a TE (Tris + EDTA) buffer solution to finally obtain a preservation solution with DNAs. The human feces DNA extraction method has the advantages of high repeatability and stability, simpleness and short time.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for extracting DNA from non-damaging sampling materials, in particular to a method for extracting human feces DNA. Background technique [0002] Colorectal cancer is the most common digestive tract malignancy, second only to gastric cancer and esophageal cancer. The incidence of colorectal cancer has gradually increased in the past two decades, and at the same time, its age of onset tends to be aging. In western developed countries, colorectal cancer is the second malignant tumor after lung cancer. Some studies in recent years have shown that mutations in some genes such as K-ras, P53, APC, and MMR can lead to the transformation of normal colonic mucosal epithelial cells into tumor cells (Vogelstein, B. and Kinzler, K.W., 1993). Studies on these gene mutations Progress has made it possible to diagnose colorectal cancer through stool DNA testing. The feces of a normal person...

AI Technical Summary

Problems solved by technology

However, there are still problems in practical application: due to the particularity of fecal materials, relevant factors such as sample collection and storage will affect the results of DNA extraction (Frantzen et al. , 1998; Piggott and Taylor, 2003)

At present, domestic and foreign literatures have not introduced these influencing factors. Therefore, it is impossible to obtain enough information from the existing literatures to guide the research on fecal DNA extraction according to the methods provided in published papers at home and abroad. less than expected effect