Method of extracting nucleic acids and method of detecting speci

Details Method of extracting nucleic acids and method of detecting speci Method of extracting nucleic acids and method of detecting speci

1996-11-12

2000-03-28

Arthur, Lisa B.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 912, 536 254, 536 2541, 536 2542, C12Q 168, C12P 1934, C07H 2100

Patent

active

06043032&

ABSTRACT:
An improved method of extracting nucleic acids from a sample comprising mixing the sample with a carrier which is at least one member selected from the group consisting of dextran, acrylamide and carboxymethyl cellulose to form a liquid mixture; mixing said liquid mixture with reagent C to render the nucleic acids and the carrier insoluble, said reagent C containing at least one reagent A selected from the group consisting of guanidinium thiocyanate, guanidinium hydrochloride, potassium thiocyanate and sodium thiocyanate and at least one reagent B selected from the group consisting of n-propyl alcohol, isopropyl alcohol, n-butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, and tert-amyl alcohol; and separating the insolubilized nucleic acids and carrier from the liquid phase. The method is easier to use than the heretofore practiced procedures, involves a smaller number of steps, can reduce the possibility of aerosol generation, can be implemented within a short time, does not use phenol or chloroform, and yet it achieves a consistent efficiency in the extraction of nucleic acids. Also disclosed is a method of detecting a specified nucleic acid sequence.

REFERENCES:
patent: 5422241 (1995-06-01), Goldrick et al.
Analtical Biochemistry vol. 162, 1987 pp. 463-465; Bowtell `Rapid isolation of eukariotic DNA`.
Higuchi et al., Biotechnology 10:413-417(Apr. 1992).
Goblet et al., Nucleic Acids Research, 17(5):2144 (1989).
Sambrook et al., Molecular Cloning:A Laboratory Manual, 2.sup.nd Ed., Cold Spring Harbor Laboratory Press, N.Y. pp E.10-E.11(1989).

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