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一种从向日葵成熟叶片中获取高质量基因组DNA的优化微量制备方法。

An optimized mini-preparation method to obtain high-quality genomic DNA from mature leaves of sunflower.

作者信息

Li J T, Yang J, Chen D C, Zhang X L, Tang Z S

机构信息

School of Life Sciences, China West Normal University, Nanchong, Sichuan, PR China.

出版信息

Genet Mol Res. 2007 Oct 5;6(4):1064-71.

Abstract

In order to investigate the mutation characteristics and to further examine the genetic variation of mutant sunflower (Helianthus annuus) obtained in plants grown from seeds exposed to space conditions, only the mature tissues such as leaf and flower could be used for DNA extraction after mutation characteristics were confirmed. The rich contents of polysaccharides, tannins, secondary metabolites, and polyphenolics made it difficult to isolate high-quality DNA from mature leaves of sunflower according to previous reports. Based on the comparison of the differences in previously reported protocols, a modified method for the extraction of high-quality DNA was developed. Using this protocol, the DNA isolated from sunflower was high in quality and suitable for restriction digestion (EcoRI, HindII, BamHI), random amplified polymorphic DNA study and further molecular research. Therefore, the modified protocol was suitable for investigating the genetic variation of sunflower using mature leaf DNA.

摘要

为了研究突变特征,并进一步检测从经太空条件处理的种子长成的突变向日葵(Helianthus annuus)植株中的遗传变异,在确认突变特征后,仅可使用叶片和花朵等成熟组织进行DNA提取。根据先前的报道,向日葵成熟叶片中多糖、单宁、次生代谢产物和多酚类物质含量丰富,难以从中分离出高质量的DNA。基于对先前报道的提取方法差异的比较,开发了一种改良的高质量DNA提取方法。使用该方法从向日葵中分离出的DNA质量高,适用于限制性酶切(EcoRI、HindII、BamHI)、随机扩增多态性DNA研究及进一步的分子研究。因此,该改良方法适用于利用成熟叶片DNA研究向日葵的遗传变异。

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