Method for identifying purity of tobacco variety Zhongyan 90 by using specific molecular marker method
A technology of molecular markers and variety purity, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of narrow genetic background, limited genetic information, and limited polymorphism of flue-cured tobacco varieties, and achieve intuitive detection results , high detection efficiency and simple operation
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[0023] Example 1 Determining the characteristic band of China Tobacco 90
[0024] 1. The test materials are shown in Table 1:
[0025] Table 1 Test materials
[0026]
[0027]
[0028] 2. Sow the seeds of various test materials to be determined in Table 1 on the seedling tray in the light incubator, and wait for the plants to grow until the genomic DNA can be directly extracted.
[0029] 3. Extract the genomic DNA of the tobacco variety to be tested (using the CTAB / NaCl method):
[0030] Take about 0.2g of a young leaf of each test material, put it in a mortar, pour liquid nitrogen into it and quickly grind it into powder, and put it into 1.5ml of 0.7ml 65°C preheated CTAB extraction buffer. In a centrifuge tube, oscillate quickly to mix well, and place in a water bath at 65°C for 1 hour, during which time shake well; place it at room temperature, add an equal volume (0.7ml) of chloroform-isoamyl alcohol (v / v 24:1), and mix gently. 4°C, 8000r, centrifuge for 10min; ta...
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