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

Quick detection kit for antibiotic lysobacter loop-mediated isothermal gene amplification, and detection method

An antibiotic lysobacteria, ring-mediated constant temperature technology, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc., to achieve the effects of low equipment requirements, short detection time, and strong specificity

Inactive Publication Date: 2017-05-31
HENAN NORMAL UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant record on the rapid detection kit and detection method of loop-mediated constant temperature gene amplification specifically for antibiotic lysobacterium

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
  • Quick detection kit for antibiotic lysobacter loop-mediated isothermal gene amplification, and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Establishment of rapid detection kit and detection method for antibiotic lysobacterium loop mediated constant temperature gene amplification

[0026] Step (1), design of primers, assembly of synthetic kits

[0027] The primer sequences determined in this embodiment for detection are as follows:

[0028] Outer primer 1: GCGAGCATGCAGCAGC,

[0029] Outer primer 2: GGCAAGGTCGCGAGGAT,

[0030] Inner primer 1: CACCGGCAAGGTCGACGAGG-CGATTCCGGCCTGGGA,

[0031] Internal primer 2: CGCGCCATGTCTTCGTGGG-GCGCAGGTCGGGTTTCA;

[0032] On this basis, a rapid detection kit for constant temperature gene amplification mediated by the antibiotic lysobacterium loop is designed, which includes the following reagents:

[0033] (1) Reaction solution 1: composed of 10mmol / L deoxynucleoside triphosphate (dNTP), 10×ThermoPol Buffer reaction buffer, 150mmol / L magnesium sulfate (MgSO 4 ), 5 mol / L betaine and sterilized double distilled water (ddH 2 O) Composition;

[0034] (2) Reaction solution...

Embodiment 2

[0047] negative control

[0048] Step (1), design of primers, assembly of synthetic kits

[0049] The sequence of primers used for detection determined in this embodiment is the same as that in Example 1.

[0050] On this basis, a rapid detection kit for constant temperature gene amplification mediated by the antibiotic lysobacterium loop is designed, which includes the following reagents:

[0051] (1) Reaction solution 1: composed of 10mmol / L deoxynucleoside triphosphate (dNTP), 10×ThermoPol Buffer reaction buffer, 150mmol / L magnesium sulfate (MgSO 4 ), 5mol / L betaine and sterilized double distilled water (ddH 2 O) Composition;

[0052] (2) Reaction solution 2: composed of 10 μmol / L outer primer 1 (F3), 10 μmol / L outer primer 2 (B3), 40 μmol / L inner primer 1 (FIP) and 40 μmol / L inner primer 2 (BIP);

[0053] (3) 8U / μL Bst DNA polymerase;

[0054] (4) Chromogen: SYBR Green I fluorescent dye with a mass concentration of 10%;

[0055] The reaction solution 1 in the above-m...

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

The invention discloses a quick detection kit for antibiotic lysobacter loop-mediated isothermal gene amplification, and a detection method. The kit carries out large-scale genome analysis by virtue of a biological information platform; according to the phenazine synthesis gene phzF of antibiotic lysobacter, four specific primers are designed; a LAMP (Loop-Mediated Isothermal Amplification) technology is combined to establish a quick, sensitive and accurate detection method by aiming at the antibiotic lysobacter, and a quick detection kit used for the method is constructed. When the quick detection kit and the detection method are used, observation and identification are carried out through naked eyes after reaction is carried out, no other analysis steps including electrophoresis and the like are required, and the quick detection kit has the advantages of short detection time, high specificity, low instrument equipment requirement, convenience in operation and the like and can be used for the quick detection of the antibiotic lysobacter.

Description

technical field [0001] The present invention relates to a kit for rapid detection of bacterial samples using loop-mediated isothermalamplification of DNA (LAMP) technology, in particular to a rapid detection reagent for loop-mediated isothermalamplification of DNA (LAMP), an antibiotic lysobacterium Box and detection method. Background technique [0002] Crop diseases are an important factor restricting agricultural production, which are mainly controlled by breeding disease-resistant varieties and chemical pesticides. However, due to the non-biological origin of chemical pesticides, it is easy for harmful organisms to develop resistance, coupled with the abuse of chemical pesticides, resulting in long-term residues of chemical pesticides in nature and the food chain, causing serious consequences such as environmental pollution, ecological balance damage, and social public nuisance. Microbial pesticides have the characteristics of being safe to non-target organisms, less to...

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): C12Q1/68C12Q1/04
CPCC12Q1/689C12Q1/6844C12Q2531/119C12Q2563/107
Inventor 徐雅梦姜晓冰姜晓杰于涛张艺鸽
Owner HENAN NORMAL UNIV
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