The invention discloses a method for extracting total
DNA (deoxyribonucleic acid) of mulberry
rhizosphere soil microorganisms by adopting a plurality of measures, which can achieve a good
cell lysis effect by utilizing PVP (
polyvinyl pyrrolidone) prewashing to remove
humic acid, the
synergy of CTAB (cetyltrimethyl
ammonium bromide),
lysozyme,
protease and SDS (
sodium dodecyl sulfate) to lyse cells and repetitively freezing and thawing the cells at the same time and utilizes PEG8000 to precipitate
DNA, so that the purity of the obtained
DNA is high. The adopted method is simple and convenient, and the DNA can be used in subsequent PCR (
Polymerase Chain Reaction) analysis and DGGE (denaturing gradient
gel electrophoresis) analysis without needing to be purified. The extracted DNA is so pure as to meet the requirement of direct subsequent
molecular biology research, an extracted DNA fragment can be used in the amplification and DGGE map analysis of 16SrDNA, moreover, the specific band
gel extraction and base sequencing of a DGGE band can be carried out, comparison with the sequence existing in an RDP (ribosomal
database project)
database can be carried out, or a new sequence probe can be established to identify unknown
bacteria, so that the composition of a microbial
community structure can be determined, and thereby laying a foundation for the future research of the
community structure diversity and the ecological functions of the mulberry
rhizosphere soil microorganisms.