ACTA AGRICULTURAE BOREALI-SINICA ›› 2017, Vol. 32 ›› Issue (6): 67-72. doi: 10.7668/hbnxb.2017.06.010

Special Issue: Chinese cabbage Biotechnology

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Study on Rapid Extraction of Genomic DNA from Chinese Cabbage

WANG Tao1, WANG Chaonan2, ZHANG Hong2, WEN Juanjuan1, ZHANG Bin2   

  1. 1. College of Life Sciences, Tianjin Normal University, Tianjin 300387, China;
    2. Tianjin Kernel Vegetable Research Institute, State Key Laboratory of Vegetable Germplasm Innovation, Key Laboratory of Tianjin Vegetable Genetic and Breeding Enterprises, Tianjin 300381, China
  • Received:2017-07-18 Published:2017-12-28

Abstract: Rapid and efficient DNA extraction is the key step in large-scale molecular breeding of crop. To construct a method for rapid extraction of genomic DNA from Chinese cabbage, the Chinese cabbage leaves were used as experimental materials, the quality of DNA extracted by the CTAB method, two-step CTAB method and four alkaline lysis methods was compared, the PCR amplification effects of DNA extracted by different methods were analyzed, and the preservation time and preservation conditions of genomic DNA extracted by different methods were compared, the optimal method was selected for the application of molecular marker-assisted selection against clubroot disease.The results showed that genomic DNA extracted by the CTAB and three kinds of alkaline lysis methods could be used as a template for PCR, and the PCR amplification products could be detected by 8% non-denaturing polyacrylamide gel electrophoresis to clear bands. Among them, the alkaline lysis method Ⅲ, not only the extraction quality was good and the extraction process was simple and fast, but also it could meet the needs of high-throughput DNA extraction of Chinese cabbage, the extracted DNA was stored at 4℃ and -20℃, and the genomic DNA stored for 30 days was used as a template for PCR amplification, the products was still detected by polyacrylamide gel electrophoresis to clear bands, it illustrates that this method has a long preservation time and it has been proved that this method has a good effect on the application of molecular marker-assisted selection against clubroot disease. Alkaline lysis method Ⅲ has significantly improved the efficiency of Chinese cabbage molecular marker-assisted selection, and can be widely used in Chinese cabbage molecular marker-assisted selection breeding.

Key words: Chinese cabbage, Genomic DNA, Rapid extraction

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Cite this article

WANG Tao, WANG Chaonan, ZHANG Hong, WEN Juanjuan, ZHANG Bin. Study on Rapid Extraction of Genomic DNA from Chinese Cabbage[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(6): 67-72. doi: 10.7668/hbnxb.2017.06.010.

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