Fluorescent SSR primer composition and application in constructing white wax new species molecular fingerprint spectrum thereof

A technology of primer combination and fingerprinting, which is applied in the field of DNA fingerprinting, can solve the problems of high cost and time-consuming, and achieve the effects of low cost, less environmental impact, and convenient operation

Active Publication Date: 2019-04-19
SHANDONG FOREST SCI RES INST +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is costly, time-consuming, and can only be identified through many pairs of primers combined with detection sites.

Method used

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  • Fluorescent SSR primer composition and application in constructing white wax new species molecular fingerprint spectrum thereof
  • Fluorescent SSR primer composition and application in constructing white wax new species molecular fingerprint spectrum thereof
  • Fluorescent SSR primer composition and application in constructing white wax new species molecular fingerprint spectrum thereof

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Embodiment 1

[0028] 1. Identification of materials

[0029]Genomic DNA was extracted from the young leaves of different varieties (species) of Ash tree. The present invention selects the excellent varieties of white wax that have passed foreign and foreign inspection (identification): Luwa No. 6, Jinjian, Luwa No. 1, Donghong Baiwa, Luwa No. 2, Hongye Baiwa, Luwa No. 5, Jinye White wax, Jass white wax, autumn purple white wax, map white wax, autumn joy white wax, weeping white wax, columnar white wax, autumn fire white wax, Qingbi, Huaxiong, Lu wax No. 3, Lu wax No. 4, Jinzhi white wax. See Table 2.

[0030] 2. Extraction of DNA

[0031] 2.1 Preheat the prepared CTAB extraction buffer in a water bath at 65°C for 30 minutes, put chloroform-isoamyl alcohol (24:1), anhydrous ethanol, and 70% ethanol in a -20°C refrigerator to pre-cool for later use. The bowl needs to be pre-cooled in advance, and then pre-cooled with a small amount of liquid nitrogen before use;

[0032] 2.2 Take 0.4-0.5g...

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Abstract

The invention discloses a fluorescent SSR primer composition and application in constructing a white wax new species molecular fingerprint spectrum thereof. The fluorescent SSR primer composition is composed of SSR primer pairs of F202/R202 and F208/R208, FAM fluorescent marks are added to 5' ends of upstream primers of each pair of primers, and forward primers are composed of TP-M13 primers carrying fluorescent marks. According to the primer composition, the primer composition is obtained using a specific fingerprint spectrum constructing a white wax new species or white wax new species molecular ID card, and 20 white wax species can be completely distinguished to overcome the prior defects of morphological and physiological identification. The primer composition can provide scientific and effective reference bases for the further application researching of a DNA fingerprint spectrum on the white wax, the evaluation of white wax germ plasm resources, and the solving of species intellectual property right disputes of the white wax.

Description

technical field [0001] The invention relates to the technical field of DNA fingerprints, in particular to a combination of fluorescent SSR primers and its application in the construction of molecular fingerprints of 20 new varieties of white wax. Background technique [0002] The genus Fraxinus belongs to the Oleceae family, Oleaceae subfamily. There are more than 70 species of Ash in the world, and more than 30 species in my country. The germplasm resources are very rich. Both seedlings and cultivated species have rich genetic diversity, which may be related to the biological characteristics of alienation pollination. The identification and evaluation of germplasm resources is one of the important links in breeding research, but the botanical classification of its species or varieties is controversial. The traditional classification and identification of white wax mainly rely on morphological markers such as samara, flower, bud, leaf and pollen grains. The difference betwee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/156
Inventor 燕丽萍吴德军王因花任飞王开芳姚俊修李庆华刘翠兰李善文束德峰张子通
Owner SHANDONG FOREST SCI RES INST
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