Procedure for the specific isolation of total DNA content of bacterial germs and a kit for this purpose

摘要

Procedure for the specific isolation of total DNA content of bacterial germs of different samples, in the course of which the cells are lysated, the DNA content of the lysate is bound selectively, it is washed and then the desalinated linear polymer nucleic acid is eluted from the binding surface in an aqueous solution. Before cell lysis the nonviable bacterial cells are separated from the viable cells on the basis of their different cell surface physical-chemical characteristics, the viable cells of the sample are kept and then lysated using a mechanical and/or enzymatic, favorably lysozyme enzymatic method. After this exclusively double-stranded DNA deriving from the lysate of viable cells is bound on a —SiO2—TiO2- matrix containing chemically activated —OH and dodecylamine groups, and after washing it, the desalinated linear polymer nucleic acid is eluted in an aqueous solution.

主权项

1. Procedure for specific isolation of total DNA content of bacterial germs of samples, in the course of which cells are lysated, the DNA content of the lysate is bound selectively to a —SiO2—TiO2-matrix of the formula SiO2—TiO—OH(—HCl)C12H25—O—NH, is washed and then a desalinated linear polymer nucleic acid is eluted from the matrix in an aqueous solution, characterised by that preliminarily nonviable bacterial cells are separated from viable cells on the basis of their different cell surface physical-chemical characteristics, their different cell surface permeability and ionic affinity, using a differently penetrating wash solution containing detergent and an electrolyte, and centrifugation, as a result of which exclusively double-stranded DNA deriving from the lysate of viable cells is bound on said matrix.