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Simple repetitive sequence primer for portunus tritubereulatus

A sequence and numbering technology, which is applied in the field of simple repeat sequence primers for Portunus trituberculatus, can solve the problems of complex operating conditions, low annealing temperature, and short primers, and achieve the goal of reducing enzyme digestion steps, high annealing temperature, and long primers Effect

Inactive Publication Date: 2007-02-07
HUAIHAI INST OF TECH
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AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a simple repeat sequence primer for Portunus trituberculatus, which can overcome the defects of the prior art that the primers are too short, the annealing temperature is low, the operating conditions are complicated, and the cost is high

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0020] 1. Screening of primers:

[0021] 1. Synthesis of 100 standard primers

[0022] 100 UBC#9 standard primers were synthesized by Shanghai Sangong.

[0023] 2. Extraction of the genome of Portunus trituberculatus

[0024] Six natural populations of Portunus trituberculatus were collected from the natural sea areas of Dalian, Dongying, Lianyungang, Zhoushan, Zhanjiang and Dongshan, Fujian. The fresh samples were brought back to the laboratory, and the genomic DNA was extracted by the standard phenol-chloroform method.

[0025] 3. PCR reaction system

[0026] PCR system: 25μl, Mg 2+ The concentration is 2.5mmol / L, the volume of 10×Buffer is 2.75μl, DNA template 50ng, Taq enzyme 1U, dNTP mixture 0.5μl; reaction conditions: first 95°C pre-denaturation for 5min, then enter the following 40 cycles of denaturation at 94°C for 45s, 45°C-53°C Touchdown programmed annealing for 45s, 72°C extension for 1.5min, then 72°C extension for 10min, and finally 4°C storage.

[0027] 4. P...

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PUM

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Abstract

the invention discloses simple repeating sequence primer of three-wart swimming crab, which comprises the following parts: CAC ACA CAC ACA CAC AG,AGA GAG AGA GAG AGA GYT,AGA GAG AGA GAG AGA GYC,AGA GAG AGA GAG AGA GYA,GAG AGA GAG AGA GAG AYT,CAC ACA CAC ACA CAC ARC,CAC ACA CAC ACA CAC ARG, GTG TGT GTG TGT GTG TYA,GTG TGT GTG TGT GTG TYC,GTG TGT GTG TGT GTG TYG.

Description

technical field [0001] The invention relates to a primer applied to germplasm evaluation and genetic structure analysis of Portunus trituberculatus. Background technique [0002] At present, the molecular markers commonly used to evaluate biological genetic diversity and germplasm identification have certain defects, such as RAPD technology (Random amplified polymorphic DNA, Random amplified polymorphic DNA), often because RAPD primers are too short by 10 The base and annealing temperature are too low, generally 36-37°C, which leads to some non-specific results of the amplified product, and its reliability and authenticity are the main defects; while AFLP technology (amplified fragment length polymorphism, Amplified fragment length polymorphism) ), and its wide application is limited due to complex operating conditions such as enzyme digestion and relatively high cost. [0003] ISSR (Inter-simple sequence repeat) is an emerging molecular marker te...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12P19/34C12N15/11
Inventor 董志国
Owner HUAIHAI INST OF TECH
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