SSR (simple sequence repeat) primer and method for watermelon hybrid variety purity identification
A technology of hybrid seed purity and watermelon variety, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of long time required for identification and large seasonal restrictions, and save the cost of identification , the effect of strong specificity and strong commercial application value
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[0040] (1) Materials and methods
[0041] 1. Plant material
[0042] In this study, watermelon hybrids Bolin, Ander, Tianfu No. 2, Huang Fenghuang and their corresponding parents were used as experimental materials, including 470 grains of Bolin hybrids, 5 grains for each parent; Ande, Tianfu No. 2, Huang Fenghuang There are 94 grains for each hybrid, and 1 grain for each parent.
[0043] 2. DNA extraction and detection
[0044] Using the modified CTAB method, the genomic DNA of watermelon parents or their hybrid offspring was extracted from fresh plant leaves about 14 days old.
[0045]① Put 30 mg of fresh leaves into a 2ml centrifuge tube, add a steel ball with a diameter of 4mm, close the lid tightly, put it in liquid nitrogen for 90 seconds, and then use a tissue grinder to grind the sample;
[0046] ② Add 800µl 2% CTAB extract to the ground sample (CTAB extract needs to be preheated in advance), and bathe in 55℃ water for 20min;
[0047] ③Centrifuge at 12000rpm for 1m...
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