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Cabbage SSR fingerprint construction method

A fingerprint and construction method technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of easy to be affected by the environment, such as workload, and achieve the effect of simple economy, convenient operation, and improved test efficiency

Active Publication Date: 2013-07-10
CROP RES INST SHANDONG ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for constructing Chinese cabbage SSR fingerprints, and to use the SSR fingerprints to identify the varieties of Chinese cabbage in view of a series of problems such as identification of Chinese cabbage varieties can only be carried out in the field, easy to be affected by the environment, and heavy workload. identification

Method used

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  • Cabbage SSR fingerprint construction method
  • Cabbage SSR fingerprint construction method
  • Cabbage SSR fingerprint construction method

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Experimental program
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Effect test

Embodiment 1

[0020] 1) DNA extraction

[0021] Table 1 50 varieties tested

[0022]

[0023]

[0024] The DNA of the tested variety was extracted by the CTAB method: take 0.2 g of the young tissue mixture, put it into a 1.5 mL centrifuge tube, and grind it in liquid nitrogen. Or grind the seeds and put them in a 1.5mL centrifuge tube. Preheat the DNA extraction solution to 65°C, add 400 μL to each tube, and mix the sample evenly. Place the centrifuge tube in a metal bath or water bath at 65°C, keep it warm for 30 minutes, and take it off. During the heat preservation process, shake the centrifuge tube to fully mix the sample and the extract. Add 400 μL of 24:1 chloroform-isoamyl alcohol (V / V) to the centrifuge tube and shake to mix. Centrifuge at 10 000g for 10min. Transfer 200 μL of the supernatant to another 1.5 mL centrifuge tube, add 400 μL of -20°C pre-cooled absolute ethanol to precipitate the DNA. Centrifuge at 10 000 g for 1 min, discard the supernatant, add 500 μL of et...

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Abstract

The present invention discloses a cabbage SSR fingerprint construction method, and belongs to the technical field of agricultural cabbage breeding and application. DNA of a variety to be tested is extracted. Twenty nine pairs of core SSR primers as shown in a sequence table SEQ ID No. 1-58 are respectively used for PCR amplification of the DNA of the variety to be tested. PCR products are detected by a denatured polyacrylamide gel electrophoresis or a five-color fluorescent capillary electrophoresis to construct cabbage variety SSR fingerprints. The invention establishes a fast and accurate, economic and simple, stable and reliable cabbage DNA fingerprint certification standard experimental system, and on this basis, develops a DNA fingerprint detection method applicable for cabbage variety identification. The present invention utilizes capillary electrophoresis methods for variety identification of cabbage, enhances the experimental efficiency in time, and has the advantages of fast speed, accuracy and easy operation, and identification results are not influenced by environments.

Description

technical field [0001] The invention relates to a method for constructing Chinese cabbage SSR fingerprints, and belongs to the technical field of agricultural Chinese cabbage breeding and application. Background technique [0002] Chinese cabbage (B. campestris L. ssp. Pekinensis (Lour). Olsson, chromosome group AA, 2n=2x=20) originated in China and is an important vegetable crop with the most Chinese characteristics, the widest distribution and the largest planting area. The planting area of ​​Chinese cabbage in China accounts for about 9% to 15% of the vegetable planting area, and it has been introduced to Japan, South Korea, North Korea and other countries. Since Chinese cabbage plays an important role in vegetable production and consumption in China and East Asian countries, identification of commercial seed varieties is particularly important. [0003] At present, the main method for identification of Chinese cabbage varieties in my country is to conduct field testing ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 李汝玉张晗段丽丽王东建孙加梅李华郑永胜王雪梅
Owner CROP RES INST SHANDONG ACAD OF AGRI SCI
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