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园艺学报 ›› 2019, Vol. 46 ›› Issue (12): 2449-2454.doi: 10.16420/j.issn.0513-353x.2019-0054

• 新技术与方法 • 上一篇    下一篇

杏黄兜兰基因组DNA提取方法比较

邵亚林1,司俊波1,常 玮2,?,石家恋2,周 翔1,丁 勇1,?   

  1. 1西南林业大学,云南省高校林木生物技术重点实验室,昆明 650224;2中国科学院昆明植物研究所资源植物与生物技术重点实验室,昆明 650204
  • 出版日期:2019-12-25 发布日期:2019-12-25
  • 基金资助:
    云南省大学生创新创业训练计划项目(201810677027);云南省高校林木生物技术重点实验室开放基金项目(51700201)

Comparison of Methods of Genomic DNA Extraction from Paphiopedilum armeniacum

SHAO Yalin1,SI Junbo1,CHANG Wei2,?,SHI Jialian2,ZHOU Xiang1,and DING Yong1,?   

  1. 1Key Laboratory of Forest Biotechnology in Yunnan,Southwest Forestry University,Kunming 650224,China;2Key Laboratory of Economic Plants and Biotechnology,Kunming Institute of Botany,Chinese Academy of Sciences,Kunming 650204,China
  • Online:2019-12-25 Published:2019-12-25

摘要: 以杏黄兜兰(Paphiopedilum armeniacum)成熟叶片为试材,采用CTAB法、改良CTAB法、高盐低pH法和SDS法提取总DNA,用琼脂糖凝胶电泳和分光光度计法分别检测所得总DNA的完整性和纯度,以期获得杏黄兜兰基因组DNA的高质量提取方法。结果表明:应用CTAB法和高盐低pH法提取的DNA降解严重、完整性差,纯度低;SDS法提取的DNA虽完整性较好,但蛋白质污染严重,纯度低;改良CTAB法获得的DNA完整性好,纯度和产率较高。

关键词: 杏黄兜兰, DNA提取, CTAB法, SDS法, 比较

Abstract: This research aimed to isolate high quality genomic DNA from the mature leaves of Paphiopedilum armeniacum using CTAB method,improved CTAB method,high salt low pH method and SDS method. The quality of genomic DNA including integrity and purity was examined through OD value and agarose gel electrophoresis. The results showed that the genomic DNA obtained from the mature leaves of P. armeniacum with CTAB method and high salt and low pH method had poor purity,poor integrity and low yields,and the DNAs were degraded seriously and polluted by proteins. The genomic DNA extracted by the SDS method was intact without degradation,but was polluted by proteins and the DNA purity was low. Fortunately,the genomic DNA obtained by improved CTAB method was intact without degradation,had higher purity and higher yield. The ratio of A260/A280 is 1.7–1.9. It was proved that proteins,polyphenols and polysaccharides rich in the leaves cells of P. armeniacum,could be removed effectively,which had high purity and quality and could be used in molecular biological research,like PCR directly.

Key words: Paphiopedilum armeniacum, DNA extraction, CTAB method, SDS method, comparison

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