The major gene tatgw‑2d controlling wheat thousand-grain weight and its marking method
A technology of major gene and labeling method, which can be used in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., and can solve the problem of unclear molecular regulation mechanism of grain weight.
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[0035] The present invention will be described in further detail below according to the drawings and embodiments.
[0036] (1) Wheat Genomic DNA Extraction
[0037] The test materials used for DNA extraction were parental Jing 411 and Hongmangchun 21, two high-low mixed pools, recombinant inbred line RIL (Jing 411 / Hongmangchun 21) F 9 Groups of families. The specific method is:
[0038] 1. Grind the wheat single grain, take 0.1g and add 0.7mL SDS extract (0.1M Tris-HCl (PH=8.5), 0.1M NaCl, 0.05M EDTA (PH=8.0), 2% SDS) at 60°C Under cracking for 45min, during which several times of shock.
[0039]2. Centrifuge at 12000 rpm at 4°C for 10 minutes.
[0040] 3. Take the supernatant and add an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1) and rotate up and down until the two phases do not separate quickly.
[0041] 4. Centrifuge for 10 minutes at 12000 rpm at 4°C.
[0042] 5. Take the supernatant and add an equal volume of phenol: chloroform: isoamyl alcohol (2...
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