Methods for extracting nucleic acids from a wide range of organisms by nonlytic permeabilization

摘要

Methods for extracting nucleic acids by heating sample cells at about 80-95 degrees C. in a permeabilization reagent containing a non-ionic detergent and a metal chelating agent. The methods of the invention release large fragments of undegraded nucleic acids without physically disrupting the entire cell wall. The nucleic acids are released into solution without bursting the cells and are suitable for research and testing without further purification. The extraction method described herein is rapid, easy to perform, and applicable to a wide variety of cells, including microorganisms. Clinical samples may be screened for the presence or absence of a microorganism by heating the sample at 80-95 degrees Celsius in the presence of a non-ionic detergent and a metal chelating agent, adding to the sample a nucleic acid probe specific to the selected microorganism, incubating the sample under conditions which allow the probe to hybridize to released nucleic acid and detecting whether any hybridized probe is present. A kit for performing the nucleic acid extraction methods disclosed herein is also described.