SSR primer pair for identifying hybrid cotton variety gossypium arboreum 1279 and product and identification method thereof
A hybrid cotton and primer pair technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve problems such as the limitation of hybrid cotton, the interference of breeder judgment, and the large impact of field environment , to achieve strong specificity, stable and reliable detection
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Embodiment 1
[0066] 1. Cotton material
[0067] Seeds of the following cotton varieties: Zhongmian 1279, Upland cotton genetic standard line TM-1, Zhongmian Institute 10, Lumianyan 28, Shumian 2, Chuanmian 239, Emian 18 and Simian 2, Zhongmian Institute 12, Zhongmian Institute 49, Xinlu Zhong 69.
[0068] 2. DNA extraction
[0069] The seeds of Zhongmian 1279 and 10 other materials used as controls were removed, and the genomic DNA was extracted by SDS method, and diluted to a final concentration of 50 ng / μL.
[0070] 3. Amplification of SSR primers
[0071] Using the genomic DNA extracted above as a template, 182 pairs of primers (7 pairs of primers per chromosome on average) were selected for SSR PCR amplification for 26 chromosomes of upland cotton. The PCR reaction system was 10 μL, the amplification mixture system was shown in Table 1, and the PCR amplification program was shown in Table 2.
[0072] Table 1 PCR amplification reaction system
[0073] Reagent
volume
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Embodiment 2
[0101]The hybrid cotton variety Zhongmian 1279 and other cotton varieties were detected according to the method in Example 1, and the results obtained were completely consistent with Example 1. It shows that the SSR primer pair provided by the present invention has good detection stability, and can effectively distinguish the hybrid cotton variety Zhongmian 1279 from other varieties.
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