Isolation of nucleic acids
AssigneeExact Sciences Corporation
InventorsJanelle J. Bruinsma, Hemanth D. Shenoi, Michael J. Domanico, James P. Light, II, Keith Kopitzke, John Zeis
Life of this patent
Estimating the term…
Abstract
Provided herein is technology relating to isolating nucleic acids. In particular, the technology relates to methods and kits for extracting nucleic acids from problematic samples such as stool.
Claims22
A method for removing an assay inhibitor from a crude sample preparation comprising a nucleic acid, the method comprising:
- a) adding insoluble polyvinylpyrrolidone to said crude sample preparation prior to isolating the nucleic acid under conditions wherein said assay inhibitor binds to said polyvinylpyrrolidone to produce a complex in a sample to be filtered;
- b) separating the complex from the crude sample preparation to produce a clarified sample preparation comprising said nucleic acid
- wherein said separating comprises placing said sample to be filtered into a spin filter, said spin filter comprising
- i) a hollow body;
- ii) a bottom end;
- and
- iii) an open top end opposite the bottom end,
- wherein the hollow body is made from a porous filtering material, preferably a polyethylene porous filtering material, and wherein the bottom end is optionally made from said porous filtering material, and
- centrifuging said spin filter, wherein during said centrifuging, a fraction of said sample to be filtered passes through a porous filtering material of said spin filter to produce a filtrate.
A system for removing an assay inhibitor from a crude sample preparation comprising a nucleic acid, the system comprising:
- a) insoluble polyvinylpyrrolidone for binding the assay inhibitor and producing a complex;
- b) functionality for separating the complex from the crude sample preparation to produce a clarified sample preparation comprising said nucleic acid
- wherein said functionality for separating the complex from the crude sample preparation comprises a spin filter comprising
- i) a hollow body;
- ii) a bottom end;
- and
- iii) an open top end opposite the bottom end,
- wherein the hollow body is made from a porous filtering material;
- and
- c) functionality for retaining a clarified sample preparation comprising the nucleic acid.
A kit for removing an assay inhibitor from a crude sample preparation comprising a nucleic acid, the kit comprising:
- a) insoluble polyvinylpyrrolidone;
- and
- b) a spin column comprising a spin filter comprising:
- i) a hollow body;
- ii) a bottom end;
- and
- iii) an open top end opposite the bottom end,
- wherein the hollow body is made from a porous filtering material.
A system for removing an assay inhibitor from a crude sample preparation comprising a nucleic acid, the system comprising:
- a) insoluble polyvinylpyrrolidone;
- and
- b) a spin column comprising a spin filter comprising
- i) a hollow body;
- ii) a bottom end;
- and
- iii) an open top end opposite the bottom end,
- wherein the hollow body is made from a porous filtering material.
Dependencies are inferred from the claim text.