Molecule marking method for identifying variety purity of insect resistance hybrid cotton Suza No.3
A technology of cultivar purity and labeling method, applied in biochemical equipment and methods, analytical materials, microbial determination/inspection, etc. The effect of speeding up the quality inspection process, efficient and accurate quality control, and high accuracy
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[0029] In the following, the present invention will be further illustrated through the SSR marker analysis examples based on PCR technology.
[0030] The experimental materials in this example are commercial seeds of insect-resistant hybrid cotton "Suza No. 3" and their parent material seeds. Cultivate the seedlings indoors on a sand bed at 25-30°C for 5-7 days, and take the fresh cotyledons.
[0031] Methods: Extract the DNA of cotton cotyledons, use SSR molecular markers to carry out PCR amplification, subject the amplified products to gel electrophoresis, count the results and screen the characteristic primers.
[0032] 1. Extraction of cotton genomic DNA
[0033] (1) Put 1 piece of fresh cotton cotyledons the size of a thumb lid into a 1.5ml centrifuge tube, add 0.6ml extraction buffer [0.35M Glucose, 0.1M Tris-HCl (pH8.0), 5mM Na-EDTA (pH8) .0), 2% PVP, 1% (V / V) β-Me], electro-rotation grinding. Centrifuge at 10,000 rpm for 10 min, discard the supernatant;
[0034] (2) Add 0....
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