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

A third-generation sequencing zooplankton genome DNA extraction method and application

A zooplankton and extraction method technology, applied in the field of genetic engineering, can solve the problems of spending huge manpower and material resources, unable to meet the requirements of building a library, etc., and achieve the effects of stable extraction product purity, simple method, and efficient removal of pigments

Active Publication Date: 2021-12-03
JINAN UNIVERSITY
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of extracting Cladocera genomic DNA with conventional kits, we found that the genomic DNA solutions of some transparent species (such as Diaphanosoma, Leptodora and Cercopagidae) contained a large amount of pigment, A260 / 230 is always low, unable to meet the requirements of downstream warehouse construction
In addition, due to the small size of Cladocera species, the body length of most species is less than 2mm, and indoor cultivation requires a lot of manpower and material resources.

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
  • A third-generation sequencing zooplankton genome DNA extraction method and application
  • A third-generation sequencing zooplankton genome DNA extraction method and application
  • A third-generation sequencing zooplankton genome DNA extraction method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] (1) Individual collection of Diaphanosoma dubium, the steps are as follows:

[0064] The daphnia spp. fuzzy uses fresh living body, and this species can achieve absolute dominance in summer (June to August) in some water bodies. Therefore, in this experiment, water samples of zooplankton were collected from a lake on a university campus in Guangzhou by using a 120 μm trawl net. In the laboratory, the water samples containing zooplankton passed through 600 μm and 250 μm mesh screens in turn. Immediately transfer the zooplankton above the 250μm mesh sieve to a 1L beaker filled with drinking purified water, in which 100u / L penicillin and 100μg / L streptomycin were added in advance to reduce pollution. After 5 to 6 hours, manually select the required fuzzy Daphnia spp. under incandescent light. In order to ensure the purity of Daphnia chevrosum, the selection was repeated 2 to 3 times, and the selected individuals were put into pure water containing the same antibiotic conc...

Embodiment 2

[0078] (1) Individual collection of three kinds of Cladocera, the steps are as follows:

[0079] Three species of Cladocera (Diaphanosoma dubium, D.dubium), Leptodora richard (L.richard) and Bythotrephes longimanus (Bythotrephes longimanus , B.longimanus)) conducted an experimental comparison; where:

[0080] The daphnia spp. fuzzy uses fresh living body, and this species can achieve absolute dominance in summer (June to August) in some water bodies. Therefore, in this experiment, water samples of zooplankton were collected from a lake on a university campus in Guangzhou by using a 120 μm trawl net. In the laboratory, the water samples containing zooplankton passed through 600 μm and 250 μm mesh screens in turn. Immediately transfer the zooplankton above the 250μm mesh sieve to a 1L beaker filled with drinking purified water, in which 100u / L penicillin and 100μg / L streptomycin were added in advance to reduce pollution. After 5 to 6 hours, manually select the required fuzzy D...

Embodiment 3

[0097] (1) Individual collection of zooplankton, the steps are as follows:

[0098] For the zooplankton, all the fresh live zooplankton collected in the water body at one time were used. In this experiment, zooplankton water samples were collected from a lake on a university campus in Guangzhou by using a 120 μm trawl net. In the laboratory, the water samples containing zooplankton were passed through a 600 μm mesh sieve to remove impurities. Immediately transfer the zooplankton in the water sample to a 1L beaker filled with drinking purified water, in which 100u / L penicillin and 100μg / L streptomycin were added in advance to reduce pollution. After 5-6 hours, the zooplankton was collected again with a 120 μm mesh sieve, and put into purified water containing the same antibiotic concentration to obtain fresh individual zooplankton.

[0099] (2) Zooplankton genome DNA extraction, the steps are as follows:

[0100] 1) Take the kit ( The buffer solution GA in the company's mar...

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 third-generation sequencing zooplankton genome DNA extraction method and application. The method comprises the following steps: (1) adding 18% to 20% (V / V) β-mercaptoethanol to buffer GA, and then adding zooplankton for grinding to obtain tissue buffer; (2) adding Add protease PK for digestion, then add RNase A, let stand, centrifuge, and take the supernatant; then add solvent, centrifuge, and take the supernatant; finally add buffer GB, mix well and add absolute ethanol to get the mixture; (3) Add the mixed solution to the adsorption column CB3, perform rinsing and elution, and obtain the zooplankton genomic DNA. The method of the present invention is simple, easy to operate, and can effectively remove pigments, and the obtained genomic DNA fragments of zooplankton are complete, high in purity, and high in concentration, which provides a guarantee for the smooth development of the third-generation sequencing of zooplankton, especially cladocera .

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to a third-generation sequencing zooplankton genome DNA extraction method and application. Background technique [0002] Cladocerans belong to an ancient group of crustaceans, according to the fossil record and DNA data. Their zoogeographical features are: some faunal complexes and taxa show bipolar distribution patterns; some species have wide ranges and others have narrow ranges; isolated populations and relatively concentrated in the temperate-subtropical regions of the two hemispheres native species. These are beneficial for us to analyze the historical reasons for the formation of cladocera fauna by comparing with plants, invertebrates and vertebrates. Therefore, the genome sequencing of certain species of Cladocera is beneficial for us to carry out their population evolution and functional gene analysis, discuss the genetic structure of the species and the genetic structur...

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): C12Q1/6806C12Q1/6869C12N15/10
CPCC12N15/1003C12Q1/6806C12Q1/6869C12Q2535/122
Inventor 韩博平刘平徐少林
Owner JINAN UNIVERSITY
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