Optimization of method for extraction of template of bacteria for randomly amplified polymorphic DNA / 军事医学科学院院刊

ABSTRACT

Objective:To obtain optimum template for randomly amplified polymorphic DNA(RAPD). Methods:Four different methods(rod winding, precipitation, boiling and lysis)for extraction of template DNA were used.The length and purity of template DNA and its RAPD profile were observed separately by agarose gel electrophoresis, spectrophotometric scan assays and RAPD reaction.Results:The template DNA (>23 kb) with high purity and the same RAPD profile with 2-4 kb DNA fragments were obtained by both rod winding and precipitation method. However,the template DNA (4 kb and 2 kb,respectively) with break and dispersion and low purity was extracted by method of boiling and lysis, and 600-2 000 bp DNA fragments were seen in the similar RAPD profile.Conclusions: Template DNA extracted by rod winding or precipitation method was optimized for RAPD.