安徽农业科学2024,Vol.52Issue(3) :91-94.DOI:10.3969/j.issn.0517-6611.2024.03.021

珍稀濒危树种金钱槭的基因组DNA提取方法研究

Study on Genomic DNA Extraction Methods for the Rare and Endangered Tree Species Dipteronia sinensis Oliv.

吴晟昊 杨丽 向松竹 胡茜茜 周志翔 郑波 施雪萍 刘梅
安徽农业科学2024,Vol.52Issue(3) :91-94.DOI:10.3969/j.issn.0517-6611.2024.03.021

珍稀濒危树种金钱槭的基因组DNA提取方法研究

Study on Genomic DNA Extraction Methods for the Rare and Endangered Tree Species Dipteronia sinensis Oliv.

吴晟昊 1杨丽 1向松竹 2胡茜茜 2周志翔 3郑波 1施雪萍 1刘梅2
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作者信息

  • 1. 华中农业大学园艺林学学院,湖北武汉 430070
  • 2. 湖北神农架林区林业科学研究院,湖北神农架 442400
  • 3. 华中农业大学园艺林学学院,湖北武汉 430070;湖北神农架林区林业科学研究院,湖北神农架 442400
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摘要

[目的]明确高效提取金钱槭基因组DNA的方法.[方法]以金钱槭幼嫩叶片为材料,采用 4 种不同方法提取金钱槭基因组DNA,并通过紫外分光光度计法、琼脂糖凝胶电泳检测、限制性内切酶酶切和扩增保守基因片段等方法比较不同DNA的提取效率.[结果]高盐低pH法因操作时间较长导致DNA损失较多,提取到的DNA虽然杂质含量较低,但浓度也较低.SDS法提取效果不佳,提取到的DNA含量少且完整性差.尿素法提取的DNA质量最差,降解严重,难以用于后续分子生物学研究.改良CTAB法可以大量提取能够用于酶切及PCR扩增的DNA.[结论]金钱槭DNA高效提取方法的确定可以为相关的分子生物学研究提供科学依据和技术参考.

Abstract

[Objective]To determine efficient method for extracting genomic DNA from D.sinensis.[Method]Four methods were used to ex-tract genomic DNA from young leaves of D.sinensis.The efficiency of genomic DNA extraction was assessed by ultraviolet spectrophotometer,agarose gel electrophoresis,restriction enzyme digestion,and amplification of conserved gene fragments.[Result]The results showed that the high-salt,low-pH method led to more DNA loss due to the extended operation time,resulting in low concentration of DNA with low impurity content.The SDS method had a poor extraction efficiency,yielding DNA with low content and poor integrity.The urea method produced the worst DNA extraction,with severe degradation that renders it unsuitable for molecular biology research.The improved CTAB method success-fully extracted a large amount of DNA suitable for enzyme digestion and PCR amplification.[Conclusion]The determination of an efficient DNA extraction method will provide scientific basis and technical references for related molecular biology research of D.sinensis.

关键词

金钱槭/DNA提取/CTAB法

Key words

Dipteronia sinensis Oliv./DNA extraction/CTAB method

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基金项目

神农架林区林业科学研究院科技计划项目(SAF202107)

出版年

2024
安徽农业科学
安徽省农业科学院

安徽农业科学

影响因子:0.413
ISSN:0517-6611
参考文献量14
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