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Multiple PCR (Polymerase Chain Reaction) primer and method for quickly identifying variety of scallops

A scallop and multiplex technology is applied in the field of multiplex PCR primers for rapid identification of four scallop species, which can solve problems such as difficulty in accurately identifying scallop species, and achieve the effect of simple operation and good repeatability.

Active Publication Date: 2014-02-26
LIAONING UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, in some cases, it is difficult to accurately identify the species of scallops by traditional methods

Method used

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  • Multiple PCR (Polymerase Chain Reaction) primer and method for quickly identifying variety of scallops
  • Multiple PCR (Polymerase Chain Reaction) primer and method for quickly identifying variety of scallops
  • Multiple PCR (Polymerase Chain Reaction) primer and method for quickly identifying variety of scallops

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 A method of fast identification scallop varieties

[0033] 1) The four scallops of the known varieties are four kinds of scallops, luxurious holes scallops, bay scallops, and shrimp scallops, respectively, the closed -shell muscle tissue of the scallop, cutting, and extracting the DNA of the sample according to the conventional phenol chloroform method.Use aseptic ultra -pure water to dissolve DNA samples, adjust the DNA concentration to 8NG / μL, and prepare sample DNA respectively, that is, obtain holes scallop sample DNA, luxurious hole scallop sample DNA, Bay scallop sample DNA, and shrimp scallop sample DNA;

[0034] 2) Multi -PCR amplification of different varieties of sample DNA: The specific steps are as follows: PCR reaction system is: 5 μL 10 × PCR Buffer, DNTP with a 4μL concentration of 2.5mm, 0.4 μL concentration of 5U / μL TAQ 59 ° C annealing 30SEC, extending 1min at 72 ° C, a total of 30 cycles; 72 ° C extension of 7min; PCR products were stored at 4 °...

Embodiment 2

[0037] Example 2 A method of fast identification scallop varieties

[0038] 1) Take the closed -shell muscle tissue of the city's scallop, cut it, extract the DNA of the sample according to the conventional phenol chlorine method, dissolve DNA samples with aseptic ultra -pure water, adjust the DNA concentration to 8ng / μL, and prepare the sample DNA;

[0039] 2) Multi -PCR amplification of the sample DNA: The specific steps are as follows: PCR reaction system is: 5 μL 10 × PCR Buffer, DNTP with a 4 μL concentration of 2.5mm, 0.4 μL concentration of 5U / μL of 5U / μL TAQ 59 ° C annealing 30SEC, extending 1min at 72 ° C, a total of 30 cycles; 72 ° C extension of 7min; PCR products were stored at 4 ° C after amplification;

[0040]3) After the PCR reaction is over, the 5 μL PCR product is mixed with a sample buffer with 1 μL.Polytestinal swimming, electrophoresis about 30min, EB staining, observing the results under ultraviolet light, agar glucose gel electrophoresis detection results fi...

Embodiment 3

[0041] Example 3 A method of fast identification scallop varieties

[0042] 1) Take the closed -shell muscle tissue of the city's scallop, cut it, extract the DNA of the sample according to the conventional phenol chlorine method, dissolve DNA samples with aseptic ultra -pure water, adjust the DNA concentration to 8ng / μL, and prepare the sample DNA;

[0043] 2) Multiple PCR amplification of the sample DNA: The specific steps are as follows: PCR reaction system is: 5 μL 10 × PCR Buffer, DNTP with a 4μL concentration of 2.5mm, 0.4 μl concentration of 5U / μL TAQ 59 ° C annealing 30SEC, extending 1min at 72 ° C, a total of 30 cycles; 72 ° C extension of 7min; PCR products were stored at 4 ° C after amplification;

[0044] 3) After the PCR reaction is over, the 5 μL PCR product is mixed with a sample buffer with 1 μL.Polytestinal swimming, electrophoresis about 30min, EB staining, observing the results under ultraviolet light, agar glucose gel electrophoresis detection results image 3 S...

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Abstract

The invention relates to a multiple PCR (Polymerase Chain Reaction) primer and a method for quickly identifying variety of scallops, belonging to the field of applied biotechnology. The multiple PCR primer for quickly identifying variety of scallops consists of a chlamys farreri primer, a chlamys nobilis primer, a bay scallop primer and a comb shell primer. The identifying method comprises the following steps: extracting total DNA (Deoxyribonucleic Acid) of a scallop genome, and carrying out multiple PCR amplification; judging the variety of the scallops to be detected according to sizes of PCR products by an agarose gel electrophoresis method, wherein the bands of the PCR products of the chlamys farreri, bay scallop, comb shell and chlamys nobilis are respectively 514bp, 367bp, 205bp and 149bp. The identification primer and multiple PCR method provided by the invention can agilely and quickly determine the variety of scallop samples. Compared with conventional detection methods, the method has the advantages of simplicity in operation, strong repeatability, low cost and the like.

Description

Technical field [0001] The present invention involves the method of identification of scallops, especially the multiple PCR primers and methods that quickly identify the four scallops varieties. Background technique [0002] Scallops, Mollusca, Lamellibraanchia, Pterimorphia, Pterioida, PECTINIDAE, are one of the major economic breeding shellfish categories in my country.The main breeding scallops of my country's coasts include: Chlamys farreri ), Luxurious holes scallops ( MIMACHLAMYS NOBILIS ), Gulf scallops ( Argopecten Irradians ) Heyi scallops ( MizuhopeCten Yesoensis To.The main farm of the Paeon scallops is the coast of the northern part of my country. The main breeding area of ​​the luxurious pore scallops is the eastern and southern coast of my country. The Gulf scallops are introduced from the United States.In recent years, with the continuous maturity of modern technology to scallop breeding technology, the scale of scallop breeding has continued to expand in the coast...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2537/143C12Q2531/113
Inventor 刘宏生艾海新张力赵健
Owner LIAONING UNIVERSITY
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