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A Rapid and Accurate Detection Method for Identifying Vaccinium Plants

A detection method and accurate technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems such as deficiencies, and achieve rich and quantitative effects

Active Publication Date: 2021-07-09
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

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  • A Rapid and Accurate Detection Method for Identifying Vaccinium Plants
  • A Rapid and Accurate Detection Method for Identifying Vaccinium Plants
  • A Rapid and Accurate Detection Method for Identifying Vaccinium Plants

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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 present invention proposes a detection method for quickly and accurately identifying plants of the genus Vaccinium, using the following primer pair for detection, and the nucleotide sequences of the primer pair are as follows: the nucleotide sequences of VcSSR11‑F and VcSSR11‑R are respectively shown in SEQ ID Shown in NO:1,2; The nucleotide sequence of VcSSR14-F and VcSSR14-R is shown in SEQ ID NO:3,4 respectively; The nucleotide sequence of VcSSR19-F and VcSSR19-R is shown in SEQ ID NO: 5 and 6; the nucleotide sequences of VcSSR28‑F and VcSSR28‑R are shown in SEQ ID NO:7 and 8 respectively; the nucleotide sequences of VcSSR33‑F and VcSSR33‑R are respectively shown in SEQ ID NO:9, 10 shown. In addition, the present invention also proposes a method for constructing an allele map of Vaccinium plants. Refer to this method to expand the allelic map to enrich the number of available allelic maps.

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 Patents(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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