A method for extracting high molecular weight genomic DNA from environmental samples

Abstract

The invention belongs to the technical field of molecular biology, and relates to a method for extracting a high molecular weight genome DNA from an environmental sample. The method of the invention comprises steps of embedding, wall breaking, digestion, electrophoresis, gel recovery and dissolving. An environmental sample is suspended by a fluid suspension, added with low melting point agarose and cooled at room temperature for solidification; a gel block is put in a wall breaking buffer, incubated, and kept vibrating horizontally; the wall breaking buffer is discharged, and a digestion buffer is added to the gel block, incubated, and kept vibrating horizontally; the digestion buffer is discharged, and the gel block is washed by a washing liquid and then washed by an electrophoretic buffer; after wall breaking and digestion, the gel block is cut into small blocks and inserted into a sample application hole of the low melting point agarose gel; gap of the sample application hole is sealed by the low melting point agarose gel, and an electrophoresis is carried out in an electrophoretic buffer; the gel containing DNA is cut off after dyeing, and the DNA is recovered through an electroelution; and the DNA obtained after the electroelution is dried and dissolved in an aseptic deionized water or a TE solution, and a dissolving solution is the extracted DNA. The method of the invention has good repeatability, and operations are simple, so as to completely satisfy molecular operations of database establish, etc.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and relates to a method for extracting genome DNA from environmental samples. Background technique [0002] Various microorganisms play an extremely important role in the cycle of material elements and life processes in various environments on the earth, but because there are a large number of uncultivated microorganisms in the environment, the research on all microorganisms in the environment is more meaningful. At present, the study of all microorganisms in the environment has gone deep to the molecular level, mainly through various metagenomic techniques. Such as molecular cloning, RFLP analysis, DGGE analysis, library construction, etc., the development of these methods and technologies have put forward higher requirements for the size of DNA fragments. Generally speaking, to construct a genomic library, the initial DNA length must be more than 40kb, otherwise there are very few eff...

AI Technical Summary

Problems solved by technology

[0004] In order to solve the deficiencies in existing environmental sample genome extraction methods, the present invention provides a method for extracting high molecular weight genomic DNA from environmental samples

Images