Method for eliminating bacterial DNA from thermally stable DNA polymerase using a DNA-digesting enzyme

摘要

Technical problem In the method of the polymerase chain reaction it is possible , owing to the DNA present in the required thermally stable enzyme due to its preparation, for there to be adverse effects on the test results in certain applications. The novel method is intended to make it possible to eliminate the DNA from the thermally stable enzyme in a simple and efficient manner. Solution of the problem The thermally stable DNA polymerase is treated with a DNA-digesting enzyme in an alkaline medium containing magnesium chloride, and subsequently the digesting enzyme is inactivated. Moreover a division of the reagents required for a polymerase chain reaction takes place so that the primers consisting of DNA are added only after the pretreatment. Areas of application The described method makes it possible to eliminate bacterial DNA from a thermally stable DNA polymerase employed for the polymerase chain reaction in a simple manner and thus makes it possible to use the polymerase chain reaction for efficient screening for unknown bacterial pathogens.