DNA fingerprint detection method for authenticity identification of cotton variety

A DNA fingerprinting and detection method technology, applied in the field of molecular biology, can solve the problems of high experimental cost, lack of systematic research on variety identification, long time consumption, etc., and achieve the effects of simple and convenient operation, stable and reliable detection results, and low cost.

Inactive Publication Date: 2010-09-08
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is a lack of systematic research on species identification using the existing molecular marker technology, and there are problems such as insufficient identification ability, many operation steps, long time-consuming, high experimental cost, etc., and cannot fully meet the needs of practical testing.

Method used

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  • DNA fingerprint detection method for authenticity identification of cotton variety
  • DNA fingerprint detection method for authenticity identification of cotton variety

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Identify whether the popular hybrid cotton variety X is the main promotion cotton hybrid Zhongmian Institute 48

[0038] The detection process is as follows:

[0039] 1. DNA extraction

[0040] Randomly pick 5 dry seeds of the variety X to be tested and 48 dry seeds of Zhongmian Institute for DNA preparation

[0041](1) Shell a single cotton seed, thoroughly pulverize it and transfer it to a 2ml centrifuge tube.

[0042] (2) Add 800ul DNA extraction solution (1% SDS, 0.01mol / L EDTA 8.0, 0.7mol / L NaCl, 0.05mol / L Tris-HCl, 0.5% sorbitol, 1% PVP, 1% β-mercaptoethanol) , vortex until fully mixed, then bathe in 65°C water for 30 minutes, and shake gently at intervals of about 10 minutes.

[0043] (3) After the water bath, add an equal volume of 800 μl of a mixture of phenol, chloroform and isoamyl alcohol (the volume ratio is 25:24:1 in sequence), mix up and down until no layers are formed, and centrifuge at 10,000 rpm for 10 min.

[0044] (4) Transfer the supernatant to...

Embodiment 2

[0077] To identify whether the parent material A being used by a certain unit is the conventional cotton variety Zhongmian Institute 41 that has applied for protection, the identification operation process is as follows:

[0078] According to the steps of Example 1, 5 dried seeds of the variety A to be tested and the control variety Zhongmiansuo 41 were randomly selected for DNA preparation, PCR amplification, electrophoresis detection and silver staining.

[0079] Result analysis: Through the above tests, the DNA fingerprints of the two varieties at 26 sites were obtained. After comparison, it was found that the fingerprints of the two varieties at the 26 primer sites were completely the same. According to the judgment standard, the parental material A was Zhongmian Institute 41, a conventional cotton variety that has applied for protection.

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Abstract

The invention relates to a DNA fingerprint detection method for authenticity identification of a cotton variety. The method adopts a single cotton variety to extract genome DNA of a sample to be tested, saves time and labor and simply and rapidly obtains high-quality DNA. The method uses 26 pairs of SSR core primers to combine to carry out DNA fingerprint detection of the cotton variety authenticity. The core primers have the characteristics of high polymorphism and good stability and repeatability and are uniformly distributed on 26 chromosomes of the cotton genome. The method of the invention has simple operation, is accurate and stable and is suitable for rapid detection of cotton variety authenticity.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a DNA fingerprint detection method for the authenticity of cotton varieties. Background technique [0002] With the increasing number of cotton varieties and the increasingly narrow genetic basis, it is becoming more and more difficult to distinguish cotton varieties based on morphological traits, which brings different degrees of difficulties to various links such as seed production, operation, management, and rights protection. Such as the phenomenon of many varieties, chaos and miscellaneous. In recent years, the development of molecular biology has brought the identification of varieties to the gene level. Compared with traditional morphological methods and protein electrophoresis techniques, DNA marker technology can reveal more polymorphisms. It is accurate, reliable, simple, fast, and easy to automate. The advantage is the development trend of variety identification techn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 匡猛杨伟华许红霞王延琴周大云冯新爱
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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