Development of High Efficiency Concentration Method of DNA from River Water by Combined Process of Coagulation and Form Separation

摘要

To obtain the bacterial information on water bodies as accurate and comprehensive, it is necessary to collect the bacteria form water samples at very low concentration, and to obtain the requirement of DNA for the genetic analysis. Thus, it is necessary to develop a technology for efficiency bacterial DNA concentration from water bodies. In this study, the both E. coli as Gram-negative and Enterococcus as Gram-positive bacteria were collected from the bacterial polluted water to the foam in markedly high concentration by coagulation and foam separation with in 7 min. These bacterial DNA recovered in the 100 μL of extract more than 87.5% recovery from the foam using a kit. To test this method for three actual river waters, the amounts of 0.67 to 2.70 μg DNA were separated and concentrated in 100 μL of extract from the 1000 mL volume of the samples without obstruction of suspended substances. When the DNA extract apply to a 16S rRNA-gene sequencing analysis, it was easy to obtain the information of bacteria flora in the river. The DNA extract concentrated by the combined process of coagulation and form separation will be used to various gene analysis.