KASP primer combination for constructing DNA finger print library of grape and application

A technology of DNA fingerprinting and primer combination, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., to achieve high accuracy and resolution

Active Publication Date: 2021-04-02
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The object of the present invention is to provide a kind of KASP primer combination and application for the

Method used

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  • KASP primer combination for constructing DNA finger print library of grape and application
  • KASP primer combination for constructing DNA finger print library of grape and application
  • KASP primer combination for constructing DNA finger print library of grape and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] This embodiment provides a KASP primer combination for the construction of a grape DNA fingerprint library. The primer combination sequence is SEQ ID NO: 1 to SEQ ID NO: 138, and its specific position and sequence composition are shown in the following table:

[0045]

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052] The primer names in the above table consist of "the specific position corresponding to the chromosome number of the VIT_SNP_SNP".

Embodiment 2

[0054] The present embodiment provides a grape DNA fingerprint library comprising the KASP primer combination described in Example 1 for the construction of the grape DNA fingerprint library, and the method for constructing the fingerprint library includes the following steps:

[0055] S1, extracting the genome DNA of all experimental varieties of grapes, a total of 304 kinds in this embodiment;

[0056] S2, use gene sequencing technology to sequence the extracted genomic DNA, and at the same time use SNP marker method for gene screening to screen out high-quality SNP sites;

[0057] S3, designing and synthesizing a KASP primer combination according to the screened high-quality SNP sites, and performing PCR amplification with the designed KASP primer combination;

[0058] S4, detecting the PCR amplification product.

[0059] Among them, for all experimental varieties of grapes, 348 grape germplasms were used in this embodiment, specifically 215 Eurasian species, 95 European a...

Embodiment 3

[0088] The present embodiment provides a kind of method of grape variety identification, comprises the following steps:

[0089] A1, obtaining the DNA of the grapes to be tested;

[0090] A2, determine the genes of 46 SNP sites in the obtained DNA, compare with all the grape variety genes in the fingerprint library described in Example 2;

[0091] A3, if the fingerprint of the grape to be tested is inconsistent with the comparison result of any grape in the grape fingerprint, the grape to be tested is a new variety, and the new variety is added to the fingerprint library; The fingerprint of the tested grape is completely consistent with the comparison result of a certain kind of grape in the described grape fingerprint, and further comparison is required to determine whether the grape to be tested is the grape of this variety;

[0092] The further comparison process includes comparing the place of origin, appearance and taste.

[0093] In order to prove the identification ab...

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Abstract

The invention discloses a KASP primer combination for constructing a DNA finger print library of grape and application of KASP primer combination and relates to the technical field of finger prints. The primer combination is obtained through picking 46 grape core SNP markers on the basis of combining a bioinformatics method and scientific algorithms such as mathematical statistics and carrying outdesigning, the DNA finger print library is constructed by using this SNP marker combination so as to carry out cluster analysis and variety identification on grape varieties, thus, the detection result can be more accurate and efficient; and 333 parts of germ plasm are effectively distinguished for the first time based on this primer marker combination and are the maximum population number with reference to grape identification at present. In addition, the KASP primer combination provided by the invention has the advantages that under the condition that DNA mass meets requirements of generalKASP reaction or conventional PCR, both the accuracy and resolution of detection on grape quality are very high, the detection efficiency is 10-20 times that of a SSR marker, the detection cost is equivalent to that of the SSR marker, and meanwhile, toxic chemical reagents such as acrylamide are not required to be used during detection.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a KASP primer combination and application for constructing a grape DNA fingerprint library. Background technique [0002] Grapes, as one of the most important fruit tree species in my country, can be eaten fresh, and can be used for wine making, drying, juice making and canning. By the end of 2018, the global grape planting area (including wine grapes, table grapes and dried grapes) reached 7.449 million hectares, with an output of 77.8 million tons, of which China's grape planting area (875,000 hectares) ranked second in the world. The output (11.7 million tons) ranks first in the world (http: / / www.oiv.int / en / ), and the grape industry has become an important industry in my country and a pillar industry for farmers to increase their income and get rich. However, due to the strong asexual reproduction ability of grapes, the wide range of suitable planting, and the frequent ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2600/13C12Q2531/113
Inventor 姜建福王富强刘崇怀樊秀彩张颖李民孙磊
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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