Method of isolation of dna

Details Method of isolation of dna Method of isolation of dna

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) C07H 21/04 (2006.01) C07K 14/44 (2006.01) C12N 9/22 (2006.01) C12N 15/10 (2006.01) C12P 19/34 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2115061

2115061 9303167 PCTABS00019 A rapid and efficient method of DNA isolation, storage, and cleavage that provides DNA suitable for amplification by a sequence-specific method is provided. The method for isolating biologically active DNA from a biological sample having DNA-containing structures includes: (1) contacting a biological sample containing DNA-containing structures with a lysis and storage buffer comprising a non-amphipathic chaotropic salt sufficient to lyse DNA-containing structures in the sample and a chelating agent to preserve the DNA from degradation to form a mixture of the biological sample and the lysis and storage buffer; (2) incubating the mixture formed in step (1) with a metal-containing chemical nuclease that cleaves the DNA to DNA fragments; and (3) purifying the DNA fragments. The DNA isolated can be catenated closed circular DNA, such as kinetoplast DNA of Trypanosoma cruzi. The purified DNA fragments can be used for amplification in a sequence-based DNA amplification system employing primers that hybridize to the DNA in order to determine the presence of a specific DNA sequence in the fragements. The combination of the isolation and amplification methods can be useful for the detection of parasitic, bacterial, and viral diseases by identification of DNA sequences associated with the organisms causing them.

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