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EST-SSR (simple sequence repeat) fluorescent marker primers for scapharca broughtonii population genetic analysis and application

A technology of population genetic analysis and fluorescent labeling, which is applied in the field of EST-SSR fluorescent labeling primers and applications, can solve problems such as EST-SSR fluorescent molecular labeling, and achieve simple operation, good repeatability, and accurate experimental data.

Pending Publication Date: 2020-06-02
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

[0004] At present, there are no related reports on the application of EST-SSR fluorescent molecular markers to the genetic analysis of clam populations

Method used

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  • EST-SSR (simple sequence repeat) fluorescent marker primers for scapharca broughtonii population genetic analysis and application
  • EST-SSR (simple sequence repeat) fluorescent marker primers for scapharca broughtonii population genetic analysis and application
  • EST-SSR (simple sequence repeat) fluorescent marker primers for scapharca broughtonii population genetic analysis and application

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Screening of EST-SSR Markers for Genetic Analysis of Clam Population

[0033] (1) Collect cockles with a shell length of about 3 cm from the sea area of ​​Qingdao, extract the DNA of cockles according to the conventional protocol, dilute the concentration to 50 μg / μL for use, and perform polymorphic SSR marker screening after DNA extraction;

[0034] (2) Synthesize non-fluorescent labeled primers, use the DNA in step (1) as a template, and use a 25 μL system for PCR amplification, including 0.5 U of TaqDNA polymerase 0.4 μL, 10×PCR buffer 2.5 μL, and 0.2 mM dNTP2 .5 μL, 1 μL each of 1 μM forward primer and reverse primer and 1 μL of 50 μg / μL DNA template. The PCR amplification program is: pre-denaturation at 94°C for 3min; denaturation at 94°C for 30s, annealing for 30s (see Table 1 for the optimum annealing temperature), extension at 72°C for 30s, 30 cycles; final extension at 72°C for 10min, using 8% polyacrylamide Gel electrophoresis was used for preliminary screeni...

Embodiment 2

[0039] The application of the EST-SSR fluorescently labeled primers for the genetic analysis of the clam population in the genetic analysis of the clam population comprises the following steps:

[0040] The samples were Korean wild population HY, Korean wild population HQ and its progeny HZ, and Dalian wild population DL. Extract the DNA of each group of cockles according to the routine protocol and dilute to 50 μg / μL for later use;

[0041] Synthesize EST-SSR fluorescently labeled primers for the genetic analysis of the 5 pairs of clam populations shown in Table 1, label the forward primers of the 5 pairs of primers with FAM, and amplify according to the following PCR system: The PCR reaction system is 25 μL, including 0.5 0.4 μL of U Taq DNA polymerase, 2.5 μL of 10×PCR buffer, 2.5 μL of 0.2 mM dNTP, 1 μL of 1 μM forward primer and 1 μL of reverse primer, and 1 μL of 50 μg / μL DNA template. The PCR amplification program was: pre-denaturation at 94°C for 3 min; denaturation a...

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Abstract

The invention provides EST-SSR (simple sequence repeat) fluorescent marker primers for scapharca broughtonii population genetic analysis and application and belongs to the field of molecular biologics. Sequences of the primers are shown in SEQ ID NO.1-10. The primers are adopted for genetic structure analysis on scapharca broughtonii populations, and effective tools can be provided for evaluatingand monitoring genetic structure variation of scapharca broughtonii populations in China.

Description

technical field [0001] The invention relates to molecular markers for genetic analysis of aquatic animal populations in the field of aquaculture, in particular to EST-SSR fluorescent marker primers and applications for genetic analysis of clam populations. Background technique [0002] Scapharca broughttonii, commonly known as red shell, blood shell, etc., belongs to Mollusca, Lamellibranchia, Arcoida, Arcidae, and Scapharca. It is an important large-scale economic shellfish in the Yellow Sea and Bohai Sea region of my country. Because of its high nutritional value, delicious taste, and good market prospects, it is very popular in the northern coast of my country. In recent years, cockle farming has developed rapidly in China, and the scale of farming has continued to expand. Studies have shown that there are certain differences in heat tolerance and growth traits between Chinese cockle populations and Korean cockle populations. Some domestic farmers have introduced Korean ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/156C12Q2525/151C12Q2563/107
Inventor 吴彪王文静刘志鸿孙秀俊周丽青杨爱国
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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