Unlock instant, AI-driven research and patent intelligence for your innovation.

Reagent box for accurately discriminating catfish and large-mouth catfish and discrimination method

A technology for identifying catfish and bigmouth catfish, which is applied in the field of identifying catfish and bigmouth catfish, and can solve the problems of distinction and difficulty in correct identification, etc.

Inactive Publication Date: 2009-09-23
SOUTHWEST UNIV
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at the seedling stage, generally under 250g, the two catfish are very similar in shape, and it is difficult to distinguish them correctly based on their appearance characteristics, and even experienced fish taxonomy workers are also difficult to correctly identify them

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
  • Reagent box for accurately discriminating catfish and large-mouth catfish and discrimination method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The kit for identifying catfish and largemouth catfish can be used 100 times, and its composition is as follows:

[0038] (a) DNA extraction reagents, including

[0039] Reagent 1, 40ml, its composition is: 10mmol / L Tris-HCl, pH8.0; 2mmol / L EDTA, pH8.0;

[0040] 0.4mol / L NaCl, 1% SDS;

[0041] Reagent 2, 0.8ml, its composition is: proteinase K, 20mg / mL;

[0042] Reagent 3, 30 ml, its composition is: sodium chloride, 6mol / L;

[0043] Reagent 4: 30ml isopropanol (analytical pure);

[0044] Reagent 5: 100ml 70% ethanol.

[0045] (b) polymerase chain reaction reagents: including reaction premix reagent 6 and reagent 7;

[0046] Reagent 6, containing 100 parts of the following components (the number in parentheses is the volume of 1 part, and the total amount of 100 parts is 2437.5 μl): 10 times polymerase chain reaction buffer (containing 15 mM MgCl 2 ) (2.5 μl), specific primers 10 μM NS (0.5 μl) and 10 μM NA (0.5 μl) (please biotechnology companies (such as Shanghai...

Embodiment 2

[0061] 1. Take 50 mg of fish fins, cut them into pieces, and add 400 μl of reagent 1;

[0062] 2. Add 10 μl of Reagent 2, and bathe in water at 60°C for about 2 hours until there are no particles in the solution;

[0063] 3. Add 300 μl of reagent 3, shake and mix for 30 seconds, then centrifuge at 10,000×g for 30 minutes, and transfer the supernatant to a new centrifuge tube;

[0064] 4. Add the same volume of reagent 4 as the supernatant obtained in the previous step (step 3), place at -20°C for 1 hour, then centrifuge at 10,000×g for 15 minutes, and discard the supernatant;

[0065] 5. Wash the precipitate with 1ml of Reagent 5, centrifuge at 10,000×g for 5min, dry in the air, dissolve in sterilized double distilled water, and obtain the DNA extraction solution;

[0066] 6. PCR amplification: take 1 μl of DNA as the template for PCR amplification; mix 23.875 μl of reagent 6 with 0.125 of reagent 7; melt at 94°C for 3 minutes on a thermal cycler; then denature at 94°C for 30...

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 kit for accurately identifying catfish and largemouth catfish and a detection method thereof. The kit includes DNA extraction reagents, including reagent 1, reagent 2, reagent 3, reagent 4 and reagent 5; polymerase chain reaction reagent , including reaction premix reagent 6 and reagent 7; electrophoresis and chromogenic reagents, including reagent 8, reagent 9, reagent 10 and reagent 11. The invention particularly provides a pair of specific primers NS and NA, which can accurately identify catfish and largemouth catfish by using the pair of primers for PCR amplification reaction, which is characterized by accuracy, batch size, light damage to identified fish, and will not cause the death of identified live fish.

Description

technical field [0001] The invention relates to a kit for identifying catfish (Silurus asotus) and catfish in batches accurately and in batches, and a method for using the kit to identify catfish and catfish (Silurus meridionalis). Background technique [0002] Both catfish and largemouth catfish belong to the genus Silurus in classification. Catfish is a widely distributed species in the world, while largemouth catfish is mainly distributed in the Yangtze River, Minjiang River, Xiangjiang River and other water systems in my country. Generally, the water systems where largemouth catfish is distributed are all There are catfish distribution. Adult catfish (S. asotus) are much smaller than largemouth catfish (S. meridionalis), and they can be distinguished by their size and external features. However, in the seedling stage, generally under 250g, the two catfishes are very similar in shape, and it is difficult to distinguish them correctly based on their appearance characterist...

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): C12Q1/68
Inventor 张其中
Owner SOUTHWEST UNIV