Primer sequence for detecting purity of Jinyou 303 cucumber hybrid seeds and detection method thereof
A hybrid seed purity, hybrid seed technology, applied in DNA/RNA fragmentation, recombinant DNA technology and other directions, to achieve the effect of low cost, fast and accurate identification results, and labor saving
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Embodiment 1
[0017] The method for detecting the purity of Jinyou No. 303 cucumber hybrid seeds comprises the steps:
[0018] (1) extracting the genomic DNA of the root tip of Jinyou 303 cucumber hybrid seeds germinated for 35 hours;
[0019] (2) Carry out PCR amplification: Add Jinyou 303 cucumber hybrid seed genomic DNA 10ng in the special thin-walled tube for PCR amplification, then add 15ng of the nucleotide sequence described in SEQ ID NO.1 in the sequence listing, and in the sequence listing 15ng of the nucleotide sequence described in SEQ ID NO.2, 1 times of Mg 2+ PCR buffer solution, dNTP 2mmol, Taq DNA polymerase 0.5 unit, add sterile double distilled water to 10μl; put the special thin-walled tube for PCR amplification into the PCR instrument for amplification, the amplification conditions are: 94°C pre-denaturation for 180 seconds ; Denaturation at 94°C for 30 seconds, annealing at 50°C for 30 seconds, extension at 72°C for 60 seconds, 30 cycles; extension at 72°C for 7 minutes...
Embodiment 2
[0023] The method for detecting the purity of Jinyou No. 303 cucumber hybrid seeds comprises the steps:
[0024] (1) Genomic DNA was extracted from cucumber hybrid seeds of Jinyou 303;
[0025] (2) Perform PCR amplification: add 20ng of Jinyou 303 cucumber hybrid seed genomic DNA to the thin-walled tube dedicated to PCR amplification, then add 30ng of the nucleotide sequence described in SEQ ID NO.1 in the sequence listing, and 30ng of the nucleotide sequence described in SEQ ID NO.2, 1 times of Mg 2+ PCR buffer solution, dNTP 2mmol, Taq DNA polymerase 1 unit, add sterile double distilled water to 10μl; put the special thin-walled tube for PCR amplification into the PCR instrument for amplification, the amplification conditions are: 94°C pre-denaturation for 180 seconds ; Denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, extension at 72°C for 60 seconds, 30 cycles; extension at 72°C for 7 minutes, the amplification is complete;
[0026] (3) Gel electroph...
Embodiment 3
[0029] The method for detecting the purity of Jinyou No. 303 cucumber hybrid seeds comprises the steps:
[0030] (1) Genomic DNA was extracted from cucumber hybrid seeds of Jinyou 303;
[0031] (2) Carry out PCR amplification: Add Jinyou 303 cucumber hybrid seed genomic DNA 15ng in the special thin-walled tube for PCR amplification, then add 20ng of the nucleotide sequence described in SEQ ID NO.1 in the sequence listing, and in the sequence listing 20ng of the nucleotide sequence described in SEQ ID NO.2, 1 times containing Mg 2+ PCR buffer, dNTP 2mmol, Taq DNA polymerase 0.8 unit, add sterile double distilled water to 10μl; put the thin-walled tube dedicated for PCR amplification into the PCR machine for amplification, the amplification conditions are: 94°C pre-denaturation for 180 seconds ; Denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, extension at 72°C for 60 seconds, 30 cycles; extension at 72°C for 7 minutes, the amplification is complete;
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