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Method for amplifying nucleic acid

A nucleic acid and nucleic acid molecule technology, applied in the field of nucleic acid amplification, can solve the problem that detection sensitivity may not be sufficient, and achieve the effect of improving detection sensitivity

Inactive Publication Date: 2012-07-04
TOYO SEIKAN KAISHA LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, even with the above-mentioned method, when the detection target nucleic acid contained in the sample is in a small amount, the detection sensitivity may not necessarily be sufficient.

Method used

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  • Method for amplifying nucleic acid
  • Method for amplifying nucleic acid
  • Method for amplifying nucleic acid

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Experimental program
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Effect test

Embodiment

[0092] Hereinafter, the present invention will be described more specifically by way of examples. It should be noted that the substances without the company name after the drug name are all manufactured by Wako Pure Chemical Industry Co., Ltd. In addition, unless otherwise specified, the solvent is water.

[0093] TE buffer

[0094] 4mmol / l Tris-HCl (trishydroxymethylaminomethane) hydrochloric acid

[0095] 1mmol / l ethylenediaminetetraacetic acid disodium dihydrate (EDTA)

[0096] Adjust to pH8.0

[0097] 1.5% TAE gel for electrophoresis

[0098] 1.5% agarose

[0099] 4mmol / l Tris-HCl

[0100] 1mmol / lEDTA

[0101] 1mmol / l acetic acid

[0102] LB medium

[0103] Peptone 1.0%

[0104] Yeast extract 0.5%

[0105] Sodium Chloride 1.0%

[0106] Adjust the pH to 7.0 after dissolution

[0107] [Synthesis of circular single-stranded DNA]

[0108] Enzyme preparations and kit preparations shown below were in accordance with the instructions for use unless otherwise specifi...

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Abstract

Provided is a method for amplifying a nucleic acid on the basis of a novel principle with which it is possible to simply, efficiently, and in a short time amplify a nucleic acid having a specific base sequence, and also provided is a method for detecting a nucleic acid using the aforementioned method. The method for amplifying a nucleic acid comprises: (a) obtaining a linear DNA fragment by means of a DNA polymerase extension reaction using template DNA containing the base sequence that is the subject of amplification, and a primer pair including a primer chemically modified at the 3' terminal and having a base sequence complementary to the region adjacent to the 3' terminal side of the aforementioned base sequence; and (b) performing a strand-displacement-type DNA polymerase extension reaction with the template being single-strand DNA containing the base sequence that is the subject of amplification and the replication starting point being the 3' terminal of the linear DNA fragment obtained in (a).

Description

technical field [0001] The present invention relates to a nucleic acid amplification method, particularly a nucleic acid amplification method using a combination of a chemically modified primer and circular single-stranded DNA, a method for detecting the presence or absence of a target gene using the above method, and a detection reagent used in the above method box. Background technique [0002] Currently, analysis methods and detection methods based on specific base sequences of target genes are widely used in research institutions, medical institutions, inspection institutions, and various other sites. For example, in the analysis and detection of pathogenic microorganisms and microscopic animals such as pathogenic bacteria, molds, mites and other allergens contained in samples (body fluids or cell sheets) derived from biological sources such as humans and animals, or samples derived from the environment, Even in the presence of viruses, pollen, etc., a method of detecti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12Q1/68
CPCC12Q1/6853C12Q2531/119
Inventor 远山将史
Owner TOYO SEIKAN KAISHA LTD