Primer and method for identifying germplasm of large yellow croakers and small yellow croakers

A technology of small yellow croaker and large yellow croaker, applied in biochemical equipment and methods, microbiological measurement/testing, DNA/RNA fragments, etc., can solve problems such as difficult identification and loss of external morphological characteristics

Inactive Publication Date: 2011-01-12
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, after processing, the original external morphological characteristics have been lost, which makes it difficult to distinguish between the two.

Method used

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  • Primer and method for identifying germplasm of large yellow croakers and small yellow croakers
  • Primer and method for identifying germplasm of large yellow croakers and small yellow croakers
  • Primer and method for identifying germplasm of large yellow croakers and small yellow croakers

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: A kind of identification primer of the present invention can simultaneously identify two species of large yellow croaker and small yellow croaker, and the primer sequence is:

[0022] F1: 5'-TCGGCCTCCTGGGATTTATTGTC-3';

[0023] F2: 5'-ATTATCCTGACGGGTACGCTCTAT-3';

[0024] R1: 5'-ACACGCGGGGGTCTAAC-3';

[0025] R2: 5'-ATATGCATCGGGGTAATCTGAGTA-3'.

Embodiment 2

[0026] Embodiment 2: A kind of method of the present invention utilizes above-mentioned identification primer to establish the germplasm identification of large yellow croaker and small yellow croaker, each selects 5 large yellow croakers and small yellow croakers of mature individuals, after professional morphological identification, adopt phenol / chloroform extraction Method (see Huang Peitang et al. translation "Molecular Cloning Experiment Guide" (Third Edition), Science Press, September 2002) to extract the DNA of large yellow croaker or small yellow croaker, utilize 4 kinds of primers provided in this method, carry out its genome Amplify. The PCR reaction system is 25 μL, the various components and the final concentration are Buffer 10mM, dNTP 0.2mM, Mg 2+ 1.5mM, 0.2mM for each of the 4 primers, Taq 1U, 50ng of large yellow croaker or small yellow croaker sample DNA, make up to 25μL with double distilled water, the PCR reaction program is 95°C 5min, (95°C 30sec→55°C 30sec...

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Abstract

The invention provides a primer and method for identifying germplasm of large yellow croakers and small yellow croakers. The invention is characterized in that the primer sequences are as follows: F1: 5'-TCGGCCTCCTGGGATTTATTGTC-3'; F2: 5'-ATTATCCTGACGGGTACGCTCTAT-3', R1: 5'-ACACGCGGGGGTCTAAC-3'; and R2: 5'-ATATGCATCGGGGTAATCTGAGTA-3'. The method comprises the following steps: adopting a phenol-chloroform extraction method to extract the DNA in the samples of the large yellow croakers or small yellow croakers; carrying out polymerase chain reaction (PCR): the PCR system is 25mu L, various components include Buffer, dNTP, Mg2+, four primers, Taq and suspected DNA in the samples of the large yellow croakers or small yellow croakers, and double distilled water is used for filling to 25mu L; and detecting the product by agarose gel electrophoresis after PCR, observing, photographing and recording the result and comparing the electrophoresis result with the provided standard electrophoretogram to identify the germplasm. The method is quick and accurate.

Description

technical field [0001] The invention relates to four kinds of primers, large yellow croaker and small yellow croaker, in particular to primers and identification methods for germplasm identification of large yellow croaker and small yellow croaker. Background technique [0002] Large yellow croaker (Pseudosciaena crocea (Richardson), Large Yellow Croaker) and small yellow croaker (Pseudosciaena plyactis (Bleeker), Small Yellow Croaker) belong to Perciformes (Perciformes), Percoidei (Percoidei), Sciaenidae (Sciaenidae), Yellow croaker (Larimichthys) used to occupy half of China's "four major fisheries". In the past, there were large numbers and large fish schools, forming the yellow croaker fishing season with primary economic significance. Since the end of the 1970s, due to overfishing and environmental changes, the resources of small yellow croaker have declined severely, while the resources of large yellow croaker have been nearly exhausted. Following a series of fishery ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 柳淑芳庄志猛马慧林琳戴芳群
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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