Primer and method for recognizing zoysia japonica varieties
A technology of zoysia and varieties, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc. It can solve the problems of long cycle time and test results are easily affected by environmental factors, and achieve reliable and fast Methods, improving the quality of grass variety approval work, and shortening the determination cycle
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[0028] 1) Extraction of Zoysias Genomic DNA:
[0029] Number of samples: For each variety sample, 25 populations are used as a mixed sample.
[0030] 2) Reagent preparation method:
[0031] Formula of DNA extraction solution: weigh 6.375g of D-sorbitol, 1.21g of Tris-base, EDTA-Na 2 0.168g, dilute to 100ml with sterile water, adjust the pH range to 7.5-8.25, and store at 4°C.
[0032] Nuclear lysate formula: weigh Tris-base2.42g, EDTA-Na 2 1.86g, sodium chloride 11.69g, CTAB 2g, dilute to 100ml with sterile water, adjust the pH range to 7.5-8.0, and sterilize under high temperature and high pressure.
[0033] 5% sodium lauryl sarcosinate formula: weigh 5g of sodium lauryl sarcosinate, dilute to 100ml with sterile water, filter with filter paper, and store at room temperature.
[0034] 3) The leaves of Zoysia species were used as raw materials, and the total genome DNA was extracted by CTAB method. The specific operation was as follows: 0.1 g of fresh leaf tissue was weighe...
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