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A method for quickly distinguishing grape varieties by using rapd

A variety and grape technology, which is applied in the field of quickly distinguishing grape varieties by using RAPD, can solve the problems of difficult to judge the primers for differentiating varieties, not intuitive enough, and a large amount of operation, and achieve early identification, convenient operation, and wide versatility Effect

Inactive Publication Date: 2011-12-14
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This patent uses many primers, and the amount of operation in quality identification is very large. It is not easy to judge the primers to distinguish varieties through the clustering tree, and it is not intuitive enough.

Method used

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  • A method for quickly distinguishing grape varieties by using rapd
  • A method for quickly distinguishing grape varieties by using rapd
  • A method for quickly distinguishing grape varieties by using rapd

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Embodiment Construction

[0025] The present invention will be further described below in conjunction with the accompanying drawings.

[0026] 1. Grape varieties and materials

[0027] Select 28 known public grape varieties, the variety names are shown in Table 1.

[0028] Table 128 Grape Varieties and Corresponding Numbers

[0029]

[0030]

[0031] 2. DNA extraction, PCR reaction analysis, primer screening and species identification

[0032] Using the young grape leaves of the above 28 varieties as materials, the improved CTAB method was used to extract DNA. Among them, by adding phenol and antioxidant (β-mercaptoethanol) to effectively remove protein and phenolic substances, the prepared DNA has no gel-like substances, proteins and phenolic substances, and is suitable for PCR amplification. The extracted grape young leaf genomic DNA was detected by 0.8% agarose gel electrophoresis, and the concentration and purity of the DNA solution was detected by the Bio-Photometer nucleic acid detector ...

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Abstract

The invention discloses a method for quickly distinguishing grape varieties by utilizing RAPD, and belongs to the field of molecular biology molecular markers. This method can completely distinguish 28 grape varieties through the design of 11 RAPD random primers. The primers are screened for stability and polymorphism, and the optimal annealing temperature is determined; The bands construct the map relationship of the corresponding differentiated varieties. This method is easy to operate, fast and accurate, and 7 PCRs can distinguish 28 grape varieties at the molecular level. With primers of any two species, the method can realize the early identification of seedlings and has wide versatility in other species.

Description

technical field [0001] The invention relates to the field of molecular biology molecular markers, in particular to a method for quickly distinguishing grape varieties by using RAPD. Background technique [0002] Grape not only has a long history of cultivation, but also has a wide distribution. In recent years, with the rapid development of my country's grape industry, a large number of fine varieties have been used in grape production. Therefore, establishing a rapid and reliable identification technology system for grape varieties is of great practical significance for early identification of seedlings, protection of variety rights, and distinction of varieties in production. At present, the traditional single morphological variety identification method has been difficult to effectively distinguish or identify many varieties; moreover, many grape breeding parents are increasingly concentrated on a few elite varieties or lines, making the new varieties selected have more t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 房经贵刘崇怀王晨上官凌飞宋长年吴伟民
Owner NANJING AGRICULTURAL UNIVERSITY
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