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Specific labeled primers and detection methods of Camellia oleifera varieties Changlin No. 3 and No. 21

A technology for labeling primers and Camellia oleifera seeds, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of poor repeatability, unsuitable for variety identification, complex PCR amplification map, etc., and achieve the effect of simple method

Inactive Publication Date: 2017-12-26
ZHEJIANG FORESTRY ACAD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, these molecular marker technologies use universal primers, and their PCR amplification patterns are not only complicated, but also poor in repeatability and specificity, so they are not suitable for variety identification. DNA fingerprinting can really be used for accurate and rapid identification of this variety

Method used

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  • Specific labeled primers and detection methods of Camellia oleifera varieties Changlin No. 3 and No. 21
  • Specific labeled primers and detection methods of Camellia oleifera varieties Changlin No. 3 and No. 21
  • Specific labeled primers and detection methods of Camellia oleifera varieties Changlin No. 3 and No. 21

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Experimental program
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Effect test

Embodiment 1

[0033] (1) Extraction of Camellia oleifera Genomic DNA:

[0034] Take 0.03g of the young leaves of Camellia oleifera to be tested, add liquid nitrogen and grind them thoroughly, and extract the genome DNA using a new type of rapid plant genomic DNA extraction kit (DP3111, BioTeke, Beijing BioTek Biotechnology Co., Ltd.) to extract the genome of the Camellia oleifera cultivar Crude DNA extract. The crude DNA extract was tested for integrity, purity and concentration by 1.5% agarose gel electrophoresis and a DNA / RNA UV spectrophotometer (Nanodrop Technologies, USA). OD 260 / OD 280 DNA samples >1.8 were used for subsequent PCR amplification. DNA extracts were stored in a -20°C refrigerator for later use.

[0035] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0036] The upstream primer: 5'-AATGAATGGTGCTTTGGATT-3' and the downstream primer: 5'-CAAAACCACACCTTGAGTCC-3' were synthesized by Shanghai Biotechnology Co., Ltd.

[0037] (3) PCR a...

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Abstract

The invention relates to a pair of highly specific molecular specific marker primers for Camellia oleifera varieties Changlin 3 and Changlin 21, and a method for quickly identifying the Camellia oleifera varieties Changlin 3 and Changlin 21. The primer sequences are as follows: upstream primer: 5'-AATGAATGGTGCTTTGGATT-3', downstream primer: 5'-CAAAACCACACCTTGAGTCC-3'. The molecular specific marker primer of the present invention can quickly and early identify the Camellia oleifera improved varieties Changlin Changlin 3 and No. 21, the method is simple, quick and accurate, and is an irreplaceable molecular means for distinguishing the improved Camellia oleifera varieties by appearance characteristics.

Description

(1) Technical field [0001] The invention relates to molecular specific marker primers and identification methods of improved Camellia oleifera varieties Changlin No. 3 and Changlin No. 21. (2) Background technology [0002] Camellia oleifera is the woody oil tree species with the largest planting area and wide distribution in my country. Vigorously developing the Camellia oleifera industry is of great significance to ensuring the safety of grain and oil, increasing farmers' income, promoting the comprehensive development of mountainous areas and building a new socialist countryside. Camellia oleifera industry in my country is in the ascendant with huge development potential. At present, the total area of ​​camellia oleifera forest in my country is more than 45 million mu, but the output is very low, only about 4 kilograms of camellia oil per mu. The fundamental reason is that the vast majority of Camellia oleifera species are poor in quality or severely degraded, while a l...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 李海波王丽玲徐梁魏海龙胡传久
Owner ZHEJIANG FORESTRY ACAD
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