Isolation of nucleic acids

摘要

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.

主权项

1. A method for isolating a human target nucleic acid from a stool sample for use in a nucleic acid detection reaction, the method comprising:
a) removing an assay inhibitor from said sample to produce a clarified sample preparation, said removing comprising the steps of:
a1) homogenizing said sample to produce a homogenate;a2) centrifuging said homogenate to produce a supernatant;a3) treating said supernatant with an insoluble inhibitor-adsorbing composition to bind inhibitor, if present, in an insoluble inhibitor complex; anda4) removing said insoluble inhibitor complex from said supernatant to produce a clarified sample preparation; b) capturing said target nucleic acid from said clarified sample preparation, wherein capturing said target nucleic acid comprises:
b1) exposing said clarified sample preparation to a denaturing condition to produce a denatured sample;b2) binding said target nucleic acid in said denatured sample to a target-specific capture reagent to form a capture complex;b3) isolating said capture complex from the clarified sample preparation; andb4) recovering said target nucleic acid from said capture complex in a target nucleic acid solution; and c) performing a nucleic acid detection reaction on said target nucleic acid solution wherein at least one third of the volume of said nucleic acid detection reaction is from said target nucleic acid solution.