DNA molecule identifying method for bluish dogbane and poacynum hendersonii
A technology of DNA molecules and large-leaf white hemp is applied in the field of DNA molecular identification of apocynum and large-leaf white hemp, and can solve the problems of considerable difficulty in accurate identification, no literature reports, and high repeatability.
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Embodiment 1
[0027] 47 samples of apocynum from 12 provinces including Xinjiang, Inner Mongolia, Henan, and Jilin, and 17 samples of big-leaf white hemp from three provinces including Xinjiang, Gansu, and Qinghai (all passed expert identification), after first extracting genomic DNA; Use primers SEQ ID No.1 and primers SEQ ID No.2 to carry out PCR amplification reaction on the extracted genomic DNA. The specific procedure is: 95°C pre-denaturation for 5 minutes, 94°C for 1 minute, 50-62°C for 1 minute, 72°C Extend for 1 min, and after 30 cycles, extend for 7 min at 72°C. PCR products were detected by conventional agarose gel electrophoresis, and all samples were found to have a clear single band at 220bp.
[0028] Further use primers SEQ ID No.1 and primers SEQ ID No.2 to continue the second round of PCR reaction on the extracted genomic DNA. The specific procedures are: 95°C pre-denaturation for 5 minutes, 94°C denaturation for 1 minute, 62.7°C renaturation for 1 minute, 72°C Extended at...
Embodiment 2
[0030] Four samples of two kinds of Apocynum tea and two dried leaves of Apocynum were purchased from the market respectively. Genomic DNA was extracted from each sample separately. First, use the specific primers SEQ ID No.1 and SEQ ID No.2 to carry out the first round of PCR amplification reaction. The specific reaction procedure is: pre-denaturation at 95°C for 5 min, denaturation at 94°C for 1 min, renaturation at 50-62°C for 1 min, and 72°C. Extended at ℃ for 1 min, and after 30 cycles, extended at 72°C for 7 min. After the PCR reaction was completed, conventional agarose electrophoresis was performed, and it was found that there was a clear single band at 220 bp in the electrophoresis pattern. It shows that the 4 samples to be detected are all Apocynum or Pleurotus chinensis, not other plants.
[0031]Progressively use primers SEQ ID No.1 and primers SEQ ID No.2 to continue the second round of PCR reaction on the extracted genomic DNA. The specific procedures are: pre-...
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