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Detection method for rapidly and accurately identifying ericaceae vaccinium

A detection method and accurate technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve problems such as deficiencies, and achieve the effect of abundant quantity

Active Publication Date: 2018-10-12
ZHEJIANG NORMAL UNIVERSITY
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Problems solved by technology

Although there are SSR markers for the construction of genetic maps of blueberry varieties and related species and the screening of functional genes, the EST-SSR for Vaccinium plants is seriously insufficient. As of March 26, 2018, the Vaccinium genus published by GenBank There are only 22,402 expressed sequence tags (EST), and there is no report on the application of allelic maps to identification of blueberry varieties and their relatives

Method used

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  • Detection method for rapidly and accurately identifying ericaceae vaccinium
  • Detection method for rapidly and accurately identifying ericaceae vaccinium
  • Detection method for rapidly and accurately identifying ericaceae vaccinium

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Embodiment 1

[0060] In the present invention, the allele map construction method is as follows:

[0061] 1. DNA extraction

[0062] (1) Preparation of DNA extraction solution: 2% CTAB, 0.1M Tris, 20mM EDTA, 1.4M NaCl, pH8.0; DNA dissolution buffer: 10mM Tris; 1mM EDTA; PH=8.0;

[0063] (2) Carry out the following treatment to blueberry varieties and their related species materials:

[0064] ① Add 400 μL of DNA extraction solution and 8 μL of β-mercaptoethanol to a 1.5 mL centrifuge tube, and place it in a 65°C water bath to preheat for 15 minutes;

[0065] ②Take 0.1g of fresh leaves of blueberry varieties and their close relatives, put a little polyvinylpyrrolidone K30 (PVP-K30) in a mortar, add liquid nitrogen and grind thoroughly; transfer the ground leaves to a DNA-containing Put the extract in a centrifuge tube, mix well, put it in a 65°C water bath for 30 minutes, and shake it every 10 minutes to fully lyse the mesophyll cells;

[0066] ③ After the water bath is over, take out the ...

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Abstract

The invention provides a detection method for rapidly and accurately identifying ericaceae vaccinium. Following primer pairs are used for detection. Nucleotide sequences of the primer pairs are shownas follows: the nucleotide sequences of VcSSR11-F and VcSSR11-R are shown in SEQ ID NO:1 and SEQ ID NO:2; the nucleotide sequences of VcSSR14-F and VcSSR14-R are shown in SEQ ID NO:3 and SEQ ID NO:4;the nucleotide sequences of VcSSR19-F and VcSSR19-R are shown in SEQ ID NO:5 and SEQ ID NO:6; the nucleotide sequences of VcSSR28-F and VcSSR28-R are shown in SEQ ID NO:7 and SEQ ID NO:8; the nucleotide sequences of VcSSR33-F and VcSSR33-R are shown in SEQ ID NO:9 and SEQ ID NO:10. In addition, the invention provides an establishing method of allele image patterns of the ericaceae vaccinium. By reference to the method, the allele image patterns are expanded, and the number of the available allele image patterns is increased.

Description

technical field [0001] The invention relates to the field of molecular biology DNA marker technology and application, in particular to a method for quickly and accurately identifying blueberry varieties and their close relatives by constructing an allele map through DNA marker technology. Background technique [0002] Blueberry (blueberry) originates in North America, is Rhododendron (Ericaceae) Vaccinium (Vaccinium) plant. Due to the high nutritional value of its fruit, it has been widely planted around the world since it was domesticated and cultivated in the early 20th century. my country began to introduce and cultivate blueberries in 1983. With the continuous improvement of cultivation technology, the blueberry industry has developed rapidly, forming four main production areas in Northeast China, Shandong Peninsula, Yangtze River Basin and Yunnan-Guizhou Plateau. [0003] Blueberries are divided into cultivated species and wild species. According to the size of the tre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11C12Q1/6806
CPCC12Q1/6806C12Q1/6895C12Q2531/113C12Q2565/125
Inventor 宗宇郭卫东方茜陈心怡陈文荣李永强
Owner ZHEJIANG NORMAL UNIVERSITY
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