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High-specificity molecular specific labeled primers of oil-tea camellia fine breed Changlin No. 21 and detection method thereof

A technology for marking primers and improved varieties of camellia oleifera, which is applied to biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems such as poor repeatability, unsuitable for variety identification, and complex PCR amplification maps, etc., to achieve The effect of simple method

Inactive Publication Date: 2015-10-14
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

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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  • High-specificity molecular specific labeled primers of oil-tea camellia fine breed Changlin No. 21 and detection method thereof
  • High-specificity molecular specific labeled primers of oil-tea camellia fine breed Changlin No. 21 and detection method thereof
  • High-specificity molecular specific labeled primers of oil-tea camellia fine breed Changlin No. 21 and detection method thereof

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Experimental program
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Effect test

Embodiment 1

[0030] (1) Extraction of Camellia oleifera Genomic DNA:

[0031] 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.

[0032] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0033] The upstream primer: 5′-TCACTCTTTCAGCAGCTACCCAA-3′ and the downstream primer: 5′-CCTTTCTCTATCGCTCTCAATCTCG-3′ were synthesized by Shanghai Bioengineering Technology Co., Ltd....

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Abstract

The invention relates to a pair of high-specificity molecular specific labeled primers of an oil-tea camellia fine breed Changlin No.21 and a rapid identification method of oil-tea camellia fine breed Changlin No.21. The primers have the sequences as follows: a forward primer: 5'-TCACTCTTCAGCAGCTACCCAA-3 ', and a downstream primer: 5'-CCTTCTCTATCGCTCTCAATCTCG-3'. The molecular specific labeled primers can conduct quick early identification on oil-tea camellia fine breed Changlin No.21; the method is simple, fast and accurate, and is an irreplaceable molecular tool for discrimination of oil-tea camellia fine breed by apparent characteristics.

Description

(1) Technical field [0001] The invention relates to a molecular-specific marker primer and an identification method for the improved Camellia oleifera variety Changlin 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 large number of na...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 李海波徐梁王丽玲魏海龙胡传久
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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