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

Primer group, kit and method for detecting pink paramisgurnus population

A para-Nili, primer set technology, applied in biochemical equipment and methods, recombinant DNA technology, microorganism determination/inspection, etc. Effects of cost and time required for sequencing

Pending Publication Date: 2020-11-17
HAINAN TROPICAL OCEAN UNIV
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no report on the related research on the specific amplification primers and fluorescent probes of P.

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A method for quickly and efficiently detecting populations of pink paranis cichlids, comprising the following steps:

[0029] 1) Extract the eDNA sample of the environment to be tested, the collected sample is sediment (sufficient amount, 25mL), put the eDNA sample (sediment sample) in liquid nitrogen for full grinding: put 1g of the sediment sample into the mortar , add 5-15mL of liquid nitrogen each time, and use a stick mortar to powder;

[0030] 2) Extract the DNA from the pretreated eDNA sample with a DNA extraction kit, and dissolve the extracted DNA in 100 μL of water to obtain a DNA sample solution;

[0031] 3) For the DNA sample liquid, a primer set was used to carry out qPCR reaction, and the primer set included an upstream primer SEQ ID NO.1: 5'-ACTCAAGCACAATAGTCGTAGCA-3', a downstream primer SEQ ID NO.2: 5'-TAAGCAGAGGCAGGCAGTGA-3' and a fluorescent probe Needle sequence SEQID NO.3: 5'-ROX-CATCTTCCTACTCATCCGTATAAGCCCACT-TAMRA-3';

[0032] The reaction syste...

Embodiment 2

[0036] A method for quickly and efficiently detecting populations of pink paranis cichlids, comprising the following steps:

[0037] 1) Extract the eDNA sample of the environment to be tested. The collected sample is a water sample (1L). The water sample is filtered, and the filter membrane (diameter 47mm, pore size 0.45μm) after the eDNA sample is placed in alcohol at -20°C Preserve by soaking (until eDNA extraction);

[0038] 2) Extract the DNA from the pretreated eDNA sample with a DNA extraction kit, and dissolve the extracted DNA in 100 μL of water to obtain a DNA sample solution;

[0039] 3) Perform qPCR reaction on DNA sample solution, said primer set includes upstream primer SEQID NO.1: 5'-ACTCAAGCACAATAGTCGTAGCA-3' and downstream primer SEQID NO.2: 5'-TAAGCAGAGGCAGGCAGTGA-3', fluorescent probe SEQIDNO. 3: 5'-ROX-CATCTTCCTACTCATCCGTATAAGCCCACT-TAMRA-3';

[0040] The reaction system composition of qPCR amplification is: PremixExTaq 10 μL, 10 μM upstream primer SEQIDNO...

Embodiment 3

[0044] A method for quickly and efficiently detecting populations of pink paranis cichlids, comprising the following steps:

[0045] 1) Extract the eDNA sample of the environment to be tested. The collected sample is a water sample, and the water sample is filtered, and the filter membrane after the eDNA sample is filtered is placed in alcohol and stored at -20°C;

[0046] 2) Extract the DNA from the soaked eDNA sample with a DNA extraction kit, and dissolve the extracted DNA in 100 μL of water to obtain a DNA sample solution;

[0047] 3) Amplify the DNA sample liquid with a primer set, which includes an upstream primer SEQID NO.1: 5'-ACTCAAGCACAATAGTCGTAGCA-3', a downstream primer SEQID NO.2: 5'-TAAGCAGAGGCAGGCAGTGA-3' and a fluorescent probe sequence SEQ ID NO.3: 5'-ROX-CATCTTCCTACTCATCCGTATAAGCCCACT-TAMRA-3';

[0048] The reaction system composition of qPCR amplification is: PremixExTaq 10 μL, 10 μM upstream primer SEQIDNO.1 0.5 μL, 10 μM downstream primer SEQIDNO.2 0.5 μL...

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 provides a primer group, a kit and a method for detecting a pink paramisgurnus population, and belongs to the technical field of detection of species distribution.The primer group comprises an upstream primer, a downstream primer and a fluorescent probe; the nucleotide sequence of the upstream primer is as shown in SEQ ID No.1; the nucleotide sequence of the downstream primer is as shown in SEQ ID No.2; and the nucleotide sequence of the fluorescent probe is as shown in SEQ ID No.3. The method includes the following steps: (1) collecting an environmental eDNA sample from a to-be-detected environment; (2) extracting DNA in the environmental eDNA sample to obtain a DNA sample solution; and 3) taking the DNA sample solution as a template, carrying out qPCR amplification by usingthe primer group, measuring a Ct value, and determining a result according to the Ct value. According to the method, on the premise that the environment and indigenous fish populations are not damaged, an environmental DNA technology is adopted for rapid and efficient detection of the pink paramisgurnus populations.

Description

technical field [0001] The invention belongs to the technical field of species distribution detection, and in particular relates to a primer set, a kit and a method for detecting populations of pink paranelia. Background technique [0002] The pink paraneetroplus synspilus (Hubbs, 1935), also known as the purple crater, is a fish of the order Cichliformes, the family Cichlidae, and the genus Paraneetroplus. As a well-known ornamental fish, the pink paranylic has bright colors, its head is usually purple, and its body is red; there is a clear boundary at the back of the head, and there are some black spots on the body. This fish is strong and easy to raise, has a high acceptance of bait, and has strong fecundity; it likes weakly acidic to neutral water quality, and prefers clean soft water. In addition, the pink paranylic fish is very powerful and will uproot the scenery in the aquarium; in the wild, it has a fierce temperament and devours aquatic plants. Native to southern...

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): C12Q1/6888C12Q1/6851C12N15/11
CPCC12Q1/6888C12Q1/6851C12Q2563/107C12Q2531/113
Inventor 陈治童玉和刘艳万武波
Owner HAINAN TROPICAL OCEAN 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