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

Kit and method for extracting total DNA from midrib of citrus leaves based on magnetic bead method

A magnetic bead method and kit technology, applied in the field of analytical biology, can solve problems such as not being developed, and achieve the effects of low cost, high detection sensitivity, high concentration and purity

Active Publication Date: 2021-11-30
ZHONGKAI UNIV OF AGRI & ENG
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the market mainly sells DNA extraction kits with magnetic beads for plants or animals, and a DNA extraction kit with magnetic beads for citrus Huanglongbing has not been developed yet.

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
  • Kit and method for extracting total DNA from midrib of citrus leaves based on magnetic bead method
  • Kit and method for extracting total DNA from midrib of citrus leaves based on magnetic bead method
  • Kit and method for extracting total DNA from midrib of citrus leaves based on magnetic bead method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A kit for extracting total DNA from the midrib of citrus leaves based on the magnetic bead method, comprising a lysate, a binding solution, a washing solution 1, a washing solution 2, an eluent and a magnetic bead suspension;

[0043] The lysate consists of: 100mmol / L pH=8.0Tris-HCl, 25mmol EDTA, 0.14mol / LNaCl, 3%m / v CTAB, 1%m / v potassium hydrogensulfate, 1%m / v ascorbic acid, 2 % m / v PVP-40T, 0.1% v / v β-mercaptoethanol, solvent is ddH 2 O, made into 100 mL; among them, potassium bisulfate, PVP-40T, and β-mercaptoethanol were added before use.

[0044] The preparation of the lysate is as follows: the above-mentioned Tris-HCl, EDTA, NaCl, CTAB, potassium bisulfate, ascorbic acid, PVP-40T, β-mercaptoethanol are mixed, dissolved in ddH 2 O, made into 100mL, that is, potassium bisulfate, PVP-40T, and β-mercaptoethanol were added before use.

[0045] The composition of the binding solution is: 750mmol / L NaCl, 50mmol / L pH=7.0MOPS, 0.15% v / v Triton X-100, 15% v / v isopropanol,...

Embodiment 2

[0056] A kit for extracting total DNA from the midrib of citrus leaves based on the magnetic bead method, comprising a lysate, a binding solution, a washing solution 1, a washing solution 2, an eluent and a magnetic bead suspension;

[0057] The lysate consists of: 80mmol / L pH=7.5Tris-HCl, 15mmol EDTA, 0.1mol / LNaCl, 2%m / v CTAB, 0.5%m / v potassium hydrogensulfate, 1.5%m / v ascorbic acid, 3.0 % m / v PVP-40T, 0.2% v / v β-mercaptoethanol, solvent is ddH 2 O, made into 100 mL; among them, potassium bisulfate, PVP-40T, and β-mercaptoethanol were added before use. The preparation of the lysate refers to Example 1.

[0058] The composition of the binding solution is: 600mmol / L NaCl, 60mmol / L pH=6.0MOPS, 0.15% v / v TritonX-100, 10% v / v isopropanol, and the solvent is ddH 2 O, made into 100 mL; where, isopropanol was added before use. The preparation of the binding solution refers to Example 1.

[0059] The composition of the washing solution 1 is: 6 mol / L guanidine hydrochloride, 10 mmo...

Embodiment 3

[0064] A kit for extracting total DNA from the midrib of citrus leaves based on the magnetic bead method, comprising a lysate, a binding solution, a washing solution 1, a washing solution 2, an eluent and a magnetic bead suspension;

[0065] The composition of the lysate is: 120mmol / L pH=8.5Tris-HCl, 30mmol EDTA, 0.2mol / LNaCl, 4%m / v CTAB, 1%m / v potassium hydrogensulfate, 0.5%m / v ascorbic acid, 1.0 % m / v PVP-40T, 0.1% v / v β-mercaptoethanol, solvent is ddH 2 O, made into 100 mL; among them, potassium bisulfate, PVP-40T, and β-mercaptoethanol were added before use. The preparation of the lysate refers to Example 1.

[0066] The composition of the binding solution is: 800mmol / LNaCl, 40mmol / L pH=7.0MOPS, 0.15% v / v TritonX-100, 20% v / v isopropanol, and the solvent is ddH 2 O, made into 100 mL; where, isopropanol was added before use. The preparation of the binding solution refers to Example 1.

[0067] The composition of the washing solution 1 is: 4mol / L guanidine hydrochloride,...

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 kit and an extraction method for extracting total DNA from the midrib of citrus leaves based on a magnetic bead method. The kit includes lysate, binding solution, washing solution 1, washing solution 2, eluent and magnetic bead suspension; adopt the DNA extraction kit of the present invention to extract the total genomic DNA of citrus leaves, the extracted DNA concentration and The overall level of purity is better than the commercially available Sangon Dzup (Plant) Genomic DNA Rapid Extraction Kit and Tiangen Plant DNA Extraction Kit. The invention is a DNA extraction magnetic bead method kit and an extraction method for the detection of citrus huanglongbing. The kit and the extraction method are applicable to all citrus varieties and have broad spectrum.

Description

technical field [0001] The invention relates to the technical field of analytical biology, in particular to a kit for extracting total DNA from midribs of citrus leaves based on a magnetic bead method and an extraction method thereof. Background technique [0002] Citrus huanglongbing is a devastating disease of citrus, which has caused huge losses to the world's citrus industry. So far, the prevention and control of citrus Huanglongbing at home and abroad mainly relies on the main measures such as eliminating vectors, strengthening quarantine, cultivating disease-free seedlings, and digging up diseased trees. The prevention and control measures of citrus huanglongbing need the help of precise diagnosis technology for citrus huanglongbing. PCR detection of huanglongbing pathogen "Candidatus Liberibacter spp." is currently a reliable method for the diagnosis of citrus huanglongbing, which has the advantages of rapidity, specificity, efficiency and sensitivity. [0003] At pr...

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 Patents(China)
IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/1013C12Q1/6806C12Q2563/143C12Q2563/149
Inventor 吴仲真林进添宾淑英康聪蒲新华吴泳贤
Owner ZHONGKAI UNIV OF AGRI & ENG
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