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

Primer group, kit and method for detecting astronotus ocellatus population

A detection map and primer set technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of specific amplification primers and fluorescent probes that have not been reported, and achieve fast and efficient The effect of detection, reduction of sequencing cost, and reduction of time required for sequencing

Active Publication Date: 2020-11-13
HAINAN TROPICAL OCEAN UNIV
View PDF6 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no report on the specific amplification primers and fluorescent probes of Tuli species

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 fast and efficient detection of Tuli fish populations, 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 is used to perform qPCR reaction. The primer set includes the upstream primer SEQ ID NO.1: 5'-TTGGACTTACTGCCTTAGAACT-3', the downstream primer SEQ ID NO.2: 5'-TCCTAGTATATTGGAGAAGTGGTGAAG-3' and Fluorescent probe sequence SEQ ID NO.3: 5'-FAM-CCTCCCTAACCGCTAAACAACTCAAAACCCT-TAMRA-3';

[0032] The reaction system for qPCR amplificati...

Embodiment 2

[0036] A method for fast and efficient detection of Tuli fish populations, 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 SEQ ID NO.1: 5'-TTGGACTACTTACTGCCTTAGAACT-3' and downstream primer SEQ ID NO.2: 5'-TCCTAGTATATTGGAGAAGTGGTGAAG-3', fluorescent probe Target SEQ ID NO.3: 5'-FAM-CCTCCCTAACCGCTAAACAACTCAAAACCCT-TAMRA-3';

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

Embodiment 3

[0044] A method for fast and efficient detection of Tuli fish populations, 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 upstream primer SEQ ID NO.1: 5'-TTGGACTTACTGCCTTAGAACT-3', downstream primer SEQ ID NO.2: 5'-TCCTAGTATATTGGAGAAGTGGTGAAG-3' and fluorescent Probe sequence SEQ ID NO.3: 5'-FAM-CCTCCCTAACCGCTAAACAACTCAAAACCCT-TAMRA-3';

[0048] The composition of the qPCR amplification reaction system is: PremixExTaq 10 μL, 10 μM upstream primer SEQ ID NO.1 0.4 μL, 10 μM downstream primer SEQ ID NO.2 0.4 μL, 10 μM...

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, kit and method for detecting an astronotus ocellatus population, and belongs to the technical field of species distribution detection. The primer group comprisesan 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 comprises the following steps: (1), collecting an environment eDNA sample from a to-be-detected environment; (2), extracting DNA in the environment eDNA sample to obtain a DNA sample solution; and (3), taking the DNA sample solution as a template, carrying out qPCR amplification by using the primer group, measuring a Ct value, and judging a result according to the Ct value. The method can be used for rapidly and efficiently detecting the astronotus ocellatus population by adopting an environmental DNA technology on the premise of not damaging the environment and the indigenous fish population.

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 tulich populations. Background technique [0002] Astronotus ocellatus (Agassiz, 1831) is a fish of the order Cichliformes, family Cichlidae, and genus Astronotus. The maximum body length can reach 33 cm, and the maximum weight is about 1.1 kg. The body of the fish is oval, flat on the sides, with a large head and a large mouth. Not only does it possess teeth in its jaws, but it also has a set of pharyngeal teeth. The teeth in the jaws are small and are used for grasping, while those in the throat (pharyngeal teeth) are used for manipulation and handling of prey. The base color of the body is jet black or dark brown, with irregular orange-red patches scattered on the side of the body, interspersed with red stripes, which is map-shaped and named "map fish". There is a round black spot above the b...

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
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114Y02A40/81
Inventor 王海山李由明马春来梁柳
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