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DNA fingerprint detection method of cotton variety 'Zhongmiansuo No.49'

A technology of DNA fingerprinting and cotton varieties, applied in the field of molecular biology, can solve the problems of lack of systematic research on variety identification, insufficient identification ability, and many operation steps, and achieve stable and reliable detection results, easy to promote and use at the grassroots level, and simple operation convenient effect

Inactive Publication Date: 2014-07-23
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 of cotton variety 'Zhongmiansuo No.49'
  • DNA fingerprint detection method of cotton variety 'Zhongmiansuo No.49'
  • DNA fingerprint detection method of cotton variety 'Zhongmiansuo No.49'

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 DNA fingerprint detection method of cotton variety 'Zhongmiansuo No. 49'

[0039] The purity testing of the seed samples of 'Zhongmiansuo No. 49' includes the following steps. Among them, the primer sequence information used in this method is shown in Table 1, and the reagent preparation used is shown in Table 2.

[0040] 1. DNA extraction

[0041] (1) Hull a single cotton seed, put it in a 2ml centrifuge tube, add 100 μL of ultrapure water and 1 steel ball, put it in a fully automatic sample rapid grinder, and pulverize it fully at 60 Hz for 30 s.

[0042] (2) Add 800 μL DNA extraction solution (1% SDS, 0.01M EDTA8.0, 0.7M NaCl, 0.05M Tris-HCl, 0.5% sorbitol, 1% PVP, 1% β-mercaptoethanol), vortex until fully mixed After homogeneity, put in a 65°C water bath for 30 minutes, and shake gently at intervals of about 10 minutes.

[0043] (3) After the water bath is over, add an equal volume of 800 μL of a mixture of phenol, chloroform and isoamyl alcohol (the vo...

Embodiment 2

[0080] Application of the DNA Fingerprint Detection Method of Example 2 Cotton Variety 'Zhongmiansuo No. 49'

[0081] Identify whether the conventional cotton variety X that is selling well in the market is the leading cotton variety 'Zhongmiansuo No. 49'.

[0082] According to the steps of Example 1, 24 dried seeds of the variety X to be tested and the control variety Zhongmiansuo 49 were randomly selected for DNA preparation. A total of 10 pairs of characteristic primers for BNL1154 were used for PCR amplification, electrophoresis and silver staining detection.

[0083] Result analysis: Through the above detection, the DNA fingerprints of the two varieties at 10 sites are obtained and compared. If the fingerprints of the two varieties at the 10 characteristic primer sites are exactly the same, it indicates that the X variety is a conventional cotton variety. So 49; if the two varieties have different fingerprints at more than or equal to 2 characteristic primer sites, it in...

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Abstract

The invention provides a DNA fingerprint detection method of a cotton variety 'Zhongmiansuo No.49'. The method utilizes an SSR marking technology, and adopts at least one characteristic primer selected from CIR246, DPL0917, BNL3033, BNL1053, NAU3839, HAU0878, DPL0176, NAU3424, Gh132 and BNL1154 for PCR detection. The method comprises the following steps of DNA extraction, SSR-PCR amplification, electrophoresis and silver-staining detection. The method is simple, rapid, accurate and stable, and requires only 2 d to complete purity and authenticity detection of one 'Zhongmiansuo No.49' sample.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a DNA fingerprint detection method of the cotton variety 'No. 49 Zhongmiansuo'. Background technique [0002] 'Zhongmiansuo 49' is a conventional early and mid-maturing upland cotton variety bred by the Cotton Research Institute of the Chinese Academy of Agricultural Sciences. This variety is one of the main cotton varieties planted in the Northwest inland cotton region. From 2006 to 2013, it was recommended by the Ministry of Agriculture as the leading cotton variety in Northwest China for 8 consecutive years; from 2008 to 2013, it was listed as the regional test control variety of cotton early and mid-maturity group in the inland cotton region of Northwest China; from 2007 to 2013 , has been listed as the regional test control variety of the early and mid-maturing cotton group in the autonomous region for 7 consecutive years; from 2005 to 2012, the cumulative promotion area in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2565/125C12Q2563/173
Inventor 匡猛杨伟华严根土王延琴周大云方丹马磊许红霞冯新爱
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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