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Molecular-specific marker primer and identification method for improved Camellia oleifera

A technology for marking primers and improved varieties of camellia oleifera, which is applied in the field of genetic engineering, can solve the problems of poor repeatability, complex PCR amplification patterns, and unsuitable for variety identification, and achieve the effect of simple methods

Active Publication Date: 2018-09-28
ZHEJIANG FORESTRY ACAD
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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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  • Molecular-specific marker primer and identification method for improved Camellia oleifera

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Experimental program
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Effect test

Embodiment

[0031] (1) Extraction of genomic DNA of Camellia oleifera:

[0032] Take 0.03 g of young leaves of Camellia oleifera to be tested, add liquid nitrogen and grind them thoroughly, and use a new type of rapid plant genomic DNA extraction kit (DP3111, BioTeke, Beijing BioTeke Biotechnology Co., Ltd.) Genomic DNA crude extract. The crude DNA extract was tested for integrity, purity and concentration by 1.5% agarose gel electrophoresis and 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.

[0033] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0034] The upstream primer: 5′-CGGACTGATTATCCTTGGCAAT-3′ (SEQ ID No.1) and the downstream primer: 5′-AGATCCCGCCCAAGACCTT-3′ (SEQ ID No.2), were synthesized by Shanghai Bioengineering Technology Co., Ltd.

[0035] (3) PCR amplification:

[0036] Composit...

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Abstract

The invention discloses a molecular specific marker primer and an identification method for fine varieties of camellia oleifera. Sequences of the molecular specific marker primer include upstream primers 5'-CGGACTGATTATCCTTGGCAAT-3' and downstream primers 5'-AGATCCCGCCCAAGACCTT-3'. The molecular specific marker primer and the identification method have the advantages that the fine varieties NO.21 of the camellia oleifera can be quickly identified by the molecular specific marker primer at early stages, the identification method is simple, speedy and accurate, and the molecular specific marker primer and the identification method are irreplaceable molecular means for discriminating the fine varieties of the camellia oleifera by the aid of appearance features.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a molecular-specific marker primer and an identification method for improved Camellia oleifera. Background technique [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 root cause is that most varieties of camellia oleifera are poor in quality or seriously degraded, while a large nu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 沈爱华袁位高吴初平江波李婷婷李海波
Owner ZHEJIANG FORESTRY ACAD
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