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
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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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