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Xinluo ZHANG, Jingyu PENG, Yuchang WANG, et al. Comparison of Extraction Methods of Total DNA and Optimization of SSR Reaction Systems from Docynia delavayi[J]. JOURNAL OF YUNNAN AGRICULTURAL UNIVERSITY（Natural Science）, 2022, 37(6): 1064-1070. DOI: 10.12101/j.issn.1004-390X(n).202107054
Citation: Xinluo ZHANG, Jingyu PENG, Yuchang WANG, et al. Comparison of Extraction Methods of Total DNA and Optimization of SSR Reaction Systems from Docynia delavayi[J]. JOURNAL OF YUNNAN AGRICULTURAL UNIVERSITY（Natural Science）, 2022, 37(6): 1064-1070. DOI: 10.12101/j.issn.1004-390X(n).202107054
shu

Comparison of Extraction Methods of Total DNA and Optimization of SSR Reaction Systems from Docynia delavayi

  • 1.

    College of Forestry, Southwest Forestry University, Kunming 650224, China

  • 2.

    College of Agronomy, China Agricultural University, Beijing 100000, China

More Information
  • Received Date: July 25, 2021
  • Revised Date: September 13, 2022
  • Accepted Date: September 13, 2022
  • Available Online: November 16, 2022
  • Published Date: November 29, 2022
    Graphical Abstract
    • Abstract
  • PurposeTo extract high-quality total DNA from the leaves of Docynia delavayi, establishing a stable SSR-PCR reaction system.
    MethodsUsing young leaves of D. delavayi as materials, total DNA was extracted by urea method, modified urea method, SDS method, CTAB method, MAGEN kit method, and magnetic beads kit method to select the best extraction method. The SSR-PCR reaction system was established by an orthogonal design L16(45) to optimize the factors affecting the SSR-PCR amplification system of D. delavayi.
    ResultsModified urea method was the best extraction method to extract the total DNA, and the mass concentration of total DNA, total amount, and A260/A280 extracted were 382.64 ng/μL, 38.26 μg and 1.87, respectively, and the bands of agarose gel electrophoresis were clear and bright, indicating good DNA integrity and high mass concentration. The optimized reaction system for SSR-PCR amplification was: template DNA 30 ng, Taq DNA polymerase 0.5 U, primers 1.25 μmol/L, Mg2+ 2.0 mmol/L, dNTPs 2.5 mmol/L, 10×PCR buffer 2.5 μL, and make-up water to 25 μL.
    ConclusionThe modified urea method can be used to extract high-quality total DNA from D. delavayi, and the established SSR-PCR reaction system provides technical support for genetic diversity analysis and molecular marker-assisted breeding of D. delavayi.
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    • References (47)
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