Rice seed fast germination QTL qGS11 molecular marker and application thereof
A technology of molecular markers and rice seeds, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., to achieve the effects of high selection efficiency, fast and simple identification methods, and low reliability
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Embodiment 1
[0030] (1) Materials and methods:
[0031] 1. Materials: Parental rice variety IR28 with high vigor and rice variety with low vigor Daguan rice, and BC constructed with IR28 as the recurrent parent 1 f 2 and BC 5 f 2 group.
[0032] 2. Extract individual DNA by SDS method.
[0033] 3. Molecular marker development: New primers were designed with Nipponbare and 9311 as the reference sequence, and the parental IR28 and Ozeki rice were used as templates for PCR amplification to screen for polymorphic markers.
[0034] 4. PCR reaction system: the volume is 25 microliters, including 2.5 microliters of 10×buffer, 25mM MgCl 2 1.5 microliters, 2.5 microliters of 4 pmol / microliter primer pair, 2 microliters of 2.5mM dNTPs, 0.2 microliters of 5 units / microliter Taq enzyme, 20 nanograms of template DNA, and add water to 25 microliters. The reaction program was DNA pre-denaturation at 95°C for 5 min; pre-denaturation at 95°C for 30 s, annealing at 50°C for 30 s, extension at 72°C fo...
Embodiment 2
[0043] (1) Materials and methods:
[0044] 1. Materials: rice variety IR28 and Daguan rice
[0045] 2. Extract individual DNA by SDS method.
[0046] 3. Marks: YB20, YB21 and RM26614.
[0047] 4. PCR reaction system: the volume is 25 microliters, including 2.5 microliters of 10×buffer, 25mM MgCl 2 1.5 microliters, 2.5 microliters of 4 pmol / microliter primer pair, 2 microliters of 2.5mM dNTPs, 0.2 microliters of 5 units / microliter Taq enzyme, 20 nanograms of template DNA, and add water to 25 microliters. The reaction program was DNA pre-denaturation at 95°C for 5 min; pre-denaturation at 95°C for 30 s, annealing at 50°C for 30 s, extension at 72°C for 30 s, and 35 cycles; finally, extension at 72°C for 10 min. PCR amplification was carried out on a biometer amplification instrument, and the amplified products were separated by electrophoresis on 8% non-denaturing polyacrylamide gel (containing 7.6 g of acrylamide and 0.4 g of methylenebisacrylamide in 100 ml of polyacrylami...
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