Indel Marker of Cotton Cytoplasmic Male Sterile Restorer Line and Its Identification Method
A male sterility, cytoplasmic technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of long time, poor accuracy, large consumption, etc. , easy to detect effects
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[0039] Example 1 Molecular Identification Method of Cotton Cytoplasmic Male Sterile Restorer Line
[0040] 1. DNA extraction
[0041] Referring to the mature cotton CTAB DNA extraction technology, restorer lines Zhonghui 46 (CGMCC5166, disclosed in patent application No. 201210092777.6) and Zhonghui 80 (CGMCC5167, disclosed in patent application No. Peel off the shells of 5 seeds of Cotton Institute 83 and conventional cotton varieties Zhongmiansuo 45 and Zhongmiansuo 69, grind each seed separately and transfer to a 2ml centrifuge tube; add 1000 μl DNA extraction solution (0.05M Tris-Cl , 0.01MEDTA, 2% SDS, 1% PVP, 0.5% sorbitol, 0.705M NaCl, PH = 8.0, autoclaved; add 1% β-mercaptoethanol before use), vortex and mix well, 65 ℃ water bath for 30min, Shake the centrifuge tube gently at intervals of 10 minutes; after the water bath, add 800 μl phenol: chloroform: isoamyl alcohol (25:24:1), mix up and down until there is no layering (gentle action, enough time), centrifuge at 120...
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