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Specificity labeling primer for tea-oil tree improved variety Chang Lin 18 and detection method

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

Inactive Publication Date: 2015-09-30
ZHEJIANG FORESTRY ACAD
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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 tea-oil tree improved variety Chang Lin 18 and detection method
  • Specificity labeling primer for tea-oil tree improved variety Chang Lin 18 and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] (1) Extraction of Camellia oleifera Genomic DNA:

[0039] 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.

[0040] (2) Design specific PCR amplification primers, the sequence of the primer pair is:

[0041] The upstream primer: 5'-AGCCAAAATTGCGTCAAGTCTTCT-3' and the downstream primer: 5'-AGCAAGCAAAACCACCCCCCAA-3' were synthesized by Shanghai Bioengineering Technology Co., Ltd. ...

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Abstract

The invention relates to a molecule specificity labeling primer for the high-specificity tea-oil tree improved variety Chang Lin 18 and a detection method capable of rapidly detecting tea-oil tree improved variety Chang Lin 18. According to the primer sequence, the upstream primer is 5'-AGCCAAAATTGCGTCAAGTCTTCT-3', and the downstream primer is 5'-AGCAAGCAAAACCACACCCCCAA-3'. The molecule specificity labeling primer can carry out rapid early-stage detection on tea-oil tree improved variety Chang Lin 18, and the method is easy, fast and accurate and is the irreplaceable molecular method for presenting characteristics and distinguishing tea-oil tree improved varieties.

Description

(1) Technical field [0001] The invention relates to a molecular-specific marker primer and an identification method of the improved Camellia oleifera variety Changlin 18. (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 nat...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 魏海龙李海波胡传久徐梁王丽玲
Owner ZHEJIANG FORESTRY ACAD
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