Specific labeled primer pair for rapidly identifying camellia oleifera improved variety Cenruan 2 and identifying method

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

Active Publication Date: 2017-05-17
GUANGXI FORESTRY RES INST
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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

Method used

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  • Specific labeled primer pair for rapidly identifying camellia oleifera improved variety Cenruan 2 and identifying method
  • Specific labeled primer pair for rapidly identifying camellia oleifera improved variety Cenruan 2 and identifying method
  • Specific labeled primer pair for rapidly identifying camellia oleifera improved variety Cenruan 2 and identifying method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Extraction of genomic DNA from leaves of Camellia oleifera Cenruan 2 and the control group

[0023] Take 0.3 g of the young leaves of the improved Camellia oleifera to be tested, add liquid nitrogen and grind them thoroughly, and use a new type of rapid plant genomic DNA extraction kit (DP320, TIANGEN, Tiangen Biochemical Technology Beijing Co., Ltd.) to extract the genomic DNA to obtain the improved Camellia oleifera Genomic DNA crude extract. The DNA crude extract was subjected to 1.5% agarose gel electrophoresis and a DNA / RNA UV spectrophotometer (Nanodrop Teehnologies, USA) to detect integrity, purity and concentration, and DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification . DNA extracts were stored in a -20°C refrigerator for later use.

[0024] (2) The sequence of the molecular-specific marker primer pair of Camellia oleifera Cenruan 2 (synthesized by Shanghai Bioengineering Technology Co., Ltd.)

[0025] Upstream primer: 5'-ATCATG...

Embodiment 2

[0034] (1) Extraction of genomic DNA from leaves of Camellia oleifera Cenruan 2 and the control group

[0035] Take 0.3 g of the young leaves of the improved Camellia oleifera to be tested, add liquid nitrogen and grind them thoroughly, and use a new type of rapid plant genomic DNA extraction kit (DP320, TIANGEN, Tiangen Biochemical Technology Beijing Co., Ltd.) to extract the genomic DNA to obtain the improved Camellia oleifera Genomic DNA crude extract. The DNA crude extract was subjected to 1.5% agarose gel electrophoresis and a DNA / RNA UV spectrophotometer (Nanodrop Teehnologies, USA) to detect integrity, purity and concentration, and DNA samples with OD260 / OD280>1.8 were used for subsequent PCR amplification . DNA extracts were stored in a -20°C refrigerator for later use.

[0036] (2) The sequence of the molecular-specific marker primer pair of Camellia oleifera Cenruan 2 (synthesized by Shanghai Bioengineering Technology Co., Ltd.)

[0037] Upstream primer: 5'-ATCATG...

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Abstract

The invention discloses a specific labeled primer pair for rapidly identifying a camellia oleifera improved variety Cenruan 2. The specific labeled primer pair comprises an upstream primer and a downstream primer with the base sequences shown as SEQ. ID. No.1 and SEQ. ID. No.2 in a sequence table. Meanwhile, the invention further provides a corresponding identifying method. According to the method, the specific labeled primer pair is taken to perform PCR amplification on camellia oleifera improved variety series, if a DNA band with the length of 756 bp is not shown in an electrophoresis result, the to-be-detected camellia oleifera variety is Cenruan 2, otherwise, the variety is not Cenruan 2. Tests prove that the method has specificity, is only applicable to identification of camellia oleifera improved variety Cenruan series, can perform rapid early identification on the camellia oleifera improved variety Cenruan 2, has the characteristics of being simple, fast, accurate and the like, and is an irreplaceable molecular technique for distinguishing camellia oleifera improved variety on the basis of apparent characteristics.

Description

technical field [0001] The invention belongs to the technical field of camellia oleifera identification, and in particular relates to a pair of specific marker primers and an identification method for rapid identification of Camellia oleifera Cenruan 2. Background technique [0002] Camellia oleifera (Camellia oleifera) is an important woody oil tree species in southern my country, and is known as the world's four largest woody edible oil plants together with oil palm, olive and coconut. Camellia oleifera seed oil is rich in oleic acid, linoleic acid and other unsaturated fatty acids, the content is generally more than 90%, its nutritional value and health value can be compared with olive oil, it is a veritable high-quality edible vegetable oil. Cenxi soft branch Camellia oleifera is the first improved variety of camellia oleifera in my country. It is a farm variety of common camellia oleifera. It is named for its tough and soft branches and drooping fruit. There are 9 high-...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 刘凯张乃燕王东雪江泽鹏
Owner GUANGXI FORESTRY RES INST
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