Molecular marking method for identifying variety of Suzhi No.1 hybrid zoysia
A molecular marker and variety identification technology, applied in the field of plant genetic markers and variety molecular identification, can solve the problems that developers and users are difficult to distinguish the authenticity of varieties and the influence of new varieties development and utilization, and achieve high repeatability and reliable identification Effect
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[0029] Utilize SRAP molecular marker technology to distinguish ' Su Zhi No. 1 ' hybridization Zoysia among the present invention and take as follows:
[0030] (1) Extraction of DNA and acquisition of identification markers
[0031] A new broad-spectrum plant genomic DNA rapid extraction kit (HF213) was used to extract DNA from the leaves of Zoysias hybrida 'Suzhi 1'.
[0032] Use 48 pairs of SRAP primers to amplify the extracted DNA by PCR. SRAP-PCR uses a 10 μL reaction system, which includes: 50ng template DNA, 10×PCR buffer, Mg 2+ 1.5mmol·L -1 , dNTPs200μmol·L -1, primer 0.2 μmol L -1 , Taq DNA polymerase 0.5U. The SRAP-PCR amplification program was: 94°C pre-denaturation for 4 min; 94°C denaturation for 1 min, 37°C annealing for 1 min, 72°C extension for 10 sec, 5 cycles; 94°C denaturation for 1 min, 50°C annealing for 1 min, 72°C extension for 10 sec, 35 cycles Cycle; extend at 72°C for 7 min after cycle, and store at 4°C. The amplification reaction was carried out ...
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