Method for identifying authenticity of watermelon varieties and SNP (Single Nucleotide Polymorphisms) primer combination special for method
A primer combination, primer set technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial assay/inspection, etc. question
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Embodiment 1
[0039] Embodiment 1, the acquisition of the primer combination that is used to identify the authenticity of watermelon varieties
[0040] 1. Discovery of 32 SNP sites
[0041] The present invention obtains 32 SNP sites based on the resequencing data of 17 watermelon representative resources. These 17 watermelon resources are rich in types, covering East Asian type (6), American type (5) and wild type (6), basically including the main ecological types of watermelon, in terms of growth period, fruit shape, peel thickness, flesh It has high genetic diversity in terms of agronomic traits such as color, seed size, and sugar content, and is representative of watermelon germplasm.
[0042] Specifically, the screening criteria for SNP loci are as follows: evenly selected positions throughout the genome, good polymorphism, low heterozygosity, MAF>0.3, good PCA clustering effect, high discrimination, and conservative 50bp sequences on both wings (no InDel , no SSR, no other SNP).
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Embodiment 2
[0055] The validity check of the primer combination that embodiment 2, embodiment 1 develop
[0056] The basic information of the 214 tested watermelon varieties in this example is shown in Table 3. The 214 tested watermelon varieties are all common fine varieties or some imported varieties from abroad.
[0057] Table 3. Basic information of 214 tested watermelon varieties
[0058]
[0059]
[0060]
[0061] 1. Obtaining the genomic DNA of the tested watermelon varieties
[0062] Genomic DNA of 214 leaves (true leaves of 30 seeds mixed) of 214 tested watermelon varieties were extracted by CTAB method to obtain genomic DNA of tested watermelon varieties.
[0063] The quality and concentration of the genomic DNA of the tested watermelon varieties must meet the requirements of PCR, and the standard of compliance is: agarose electrophoresis shows a single DNA band without obvious dispersion; the UV spectrophotometer Nanodrop2000 (Thermo) detects that the A260 / A280 ratio ...
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