Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method of detecting gene

A gene detection and target gene technology, which is applied in biochemical equipment and methods, microbial determination/inspection, fermentation, etc., and can solve problems such as inability to perform gene detection

Inactive Publication Date: 2003-01-01
三光纯药株式会社
View PDF2 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] As mentioned above, there are various methods for linking two oligonucleotides, but methods for detecting linked oligonucleotides such as LCR TM As represented, only the ligated oligonucleotides are captured by a special device, and genetic detection cannot be performed without removing excess oligonucleotides by B / F separation operations using washing

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method of detecting gene
  • Method of detecting gene
  • Method of detecting gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] (Example 1, Comparative Example 1 and Comparative Example 2) Ligation reaction in HCP (target gene: double strand)

[0080] 1. Purpose

[0081] Attempt to detect double-stranded target gene by ligation reaction of 1 cut HCP.

[0082] 2. Materials

[0083] 1) Regarding the target gene, a 60-base synthetic oligonucleotide derived from the Mycobacterium intracellulare 16s rRNA gene (hereinafter referred to as INT-①) and a synthetic oligonucleotide complementary to the target gene INT-① were produced (hereinafter referred to as INT-①R), the two were double-stranded, and the following experiments were performed.

[0084] 2) Preparation of probe 1 (hereinafter referred to as INT-1) having a region complementary to the Y region of the target gene INT-① and probe 2 (hereinafter referred to as INT-2) each region complementary to this probe 1 Constituted paired HCPs. Furthermore, probes obtained by cutting one of them (hereinafter respectively referred to as INT-1-A, INT-1-B,...

Embodiment 2～4 and comparative example 3

[0101] (Examples 2-4 and Comparative Example 3) Detection of SNPs by HCP ligation reaction

[0102] 1. Purpose

[0103] Discuss whether single-stranded target genes and SNPs can be detected by the ligation reaction of 1-cut HCP.

[0104] 2. Materials

[0105] In addition to the probe, ligase, ligation buffer, buffer and target gene INT-① used in Example 1, the target gene INT-①S1 (with the Y region of INT-① only 1 base different) was recreated. 1 base different from the part complementary to the 5' end of the Y region of HCP) and INT-①S2 (1 base different from the part complementary to the 3' end of the Y region of HCP).

[0106] 3. Method

[0107] a) Preparation of reaction solution

[0108] Add 0.5 μL of INT-1-A, INT-1-B, INT-2-C, and INT-2-D prepared at 100 pmol / μL respectively into 0.2 mL sterilized microtubes, and add 1 μL of ligase , culture buffer 2 μL, add 0.5 μL of the target gene prepared to 100 pmol / μL, pass DW 2 Prepare 20 μL of reaction solution respectively...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A method of detecting gene whereby a gene can be detected by forming a polymer by using the PALSAR method without capturing ligated oligonucleotides or washing excessive oligonucleotides. By using a plurality of probe pairs consisting of n (n<3) base sequence regions complementary to each other, one point in one side of the complementary domain, one point in each of the both sides thereof or two or more points therein of a target gene of the above-described probe pair are preliminarily cleaved and then a hybridization reaction, a ligation reaction and a dissociation reaction are conducted under temperature control. Thus the cleaved probes are ligated to form a complete probe. By using a method of constructing a probe polymer, a double-stranded polymer is formed and thus the above-described gene is detected.

Description

technical field [0001] The present invention relates to a method for detecting genes using a method for preparing a probe polymer using a plurality of paired probes (hereinafter, Sometimes referred to as HoneyComb Probe: HCP), make it hybridize to reach the interlaced state, thus form the method of double-strand polymer (USP6261846, Japanese Patent Application No. 2000-201687 and Japanese Patent Application No. 2000-98797, hereinafter referred to as PALSAR (Probe Alternation link self-assembly reaction) method). Background technique [0002] A ligation reaction has been developed to link two oligonucleotides by hybridizing them adjacent to two oligonucleotides that are complementary to the target gene or SNP, to detect the target gene or SNP polymorphic methods. [0003] The ligation reaction described above is broadly classified into a method using ligase and a method using a specific oligonucleotide without using an enzyme. [0004] Methods using ligase include T4 DNA l...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/34C12Q1/68C12Q1/682C12Q1/70
CPCC12Q1/682C12Q2537/143C12Q2533/107C12Q2525/131C12Q1/68
Inventor 薄井贡三塚真理波木井千雅子
Owner 三光纯药株式会社
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com