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Specificity labeling primer for camellia oleifera improved varieties changlin number three and twenty one and detection method

A technology for labeling primers and Camellia oleifera seeds, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of poor repeatability, complex PCR amplification patterns, and low specificity, etc. simple effect

Inactive Publication Date: 2015-09-30
ZHEJIANG FORESTRY ACAD
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  • Summary
  • Abstract
  • Description
  • 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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  • Specificity labeling primer for camellia oleifera improved varieties changlin number three and twenty one and detection method
  • Specificity labeling primer for camellia oleifera improved varieties changlin number three and twenty one and detection method
  • Specificity labeling primer for camellia oleifera improved varieties changlin number three and twenty one and detection method

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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 molecule specificity labeling primer for camellia oleifera improved varieties changlin number three and twenty one and a method capable of fast identifying the camellia oleifera improved varieties changlin number three and twenty one. The primer has the following sequence of an upstream primer body 5'-AATGAATGGTGCTTTGGATT-3' and a lower primer body 5'-CAAAACCACACCTTGAGTCC-3'. The molecule specificity labeling primer can fast identify the camellia oleifera improved varieties changlin number three and twenty one at the early stage, and the method is simple, fast and accurate and is an irreplaceable molecule means for identifying camellia oleifera improved varieties through the apparent 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 Applications(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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