秘鲁本土棉(Gossypium barbadense L.) DNA快速高效提取方案兰、秘鲁。

IF 2 Q3 BIOCHEMICAL RESEARCH METHODS
Luis Miguel Serquén Lopez, Herry Lloclla Gonzales, Wilmer Enrique Vidaurre Garcia, Ricardo Leonidas de Jesus Velez Chicoma, Mendoza Cornejo Greta
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引用次数: 0

摘要

从植物中高效提取高质量的DNA是分子研究的一个关键挑战,特别是在秘鲁原产于的物种中,由于存在多糖和酚类化合物等抑制剂。本研究提出了一种改进的基于ctab的二氧化硅柱方案,旨在克服这些限制,而不需要液氮或昂贵的试剂。在秘鲁兰巴耶克收集了当地的棉花样本,并使用简化的程序进行处理,以优化提取DNA的纯度和浓度。对8个有色纤维品种(奶油色、fifo色、浅棕色、深棕色、橙棕色、红色、细红色和白色)进行了评价,得到的DNA A260/A280比值为2.14 ~ 2.19,A260/A230比值为1.8 ~ 3.14;这些值比经典的CTAB法得到的值要高。利用ISSR和RAPD分子标记进行PCR扩增,得到了清晰而明确的条带模式。此外,提取的DNA适用于高级应用,如Sanger测序,可获得高质量的电泳图。结果表明,对于资源有限的实验室来说,该方案是一种高效、经济、适应性强的替代方案,可以从巴氏棉和其他植物物种中提取高质量的DNA。这种简化的方法促进了遗传和生物技术研究的发展,有助于了解秘鲁本土棉花的遗传资源并使其增值。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Rapid and Efficient DNA Extraction Protocol from Peruvian Native Cotton (<i>Gossypium barbadense</i> L.) Lambayeque, Peru.

Rapid and Efficient DNA Extraction Protocol from Peruvian Native Cotton (<i>Gossypium barbadense</i> L.) Lambayeque, Peru.

Rapid and Efficient DNA Extraction Protocol from Peruvian Native Cotton (<i>Gossypium barbadense</i> L.) Lambayeque, Peru.

Rapid and Efficient DNA Extraction Protocol from Peruvian Native Cotton (Gossypium barbadense L.) Lambayeque, Peru.

Efficient extraction of high-quality DNA from plants is a critical challenge in molecular research, especially in species such as Gossypium barbadense L., native to Peru, due to the presence of inhibitors such as polysaccharides and phenolic compounds. This study presents a modified CTAB-based protocol with silica columns that is designed to overcome these limitations without the need for liquid nitrogen or expensive reagents. Native cotton samples were collected in Lambayeque, Peru, and processed using a simplified procedure that optimizes the purity and concentration of the extracted DNA. Eight cultivars of G. barbadense L. with colored fibers (cream, fifo, light brown, dark brown, orange-brown, reddish, fine reddish, and white) were evaluated, yielding DNA with A260/A280 ratios between 2.14 and 2.19 and A260/A230 ratios between 1.8 and 3.14; these values are higher than those obtained with the classical CTAB method. DNA quality was validated by PCR amplification using ISSR and RAPD molecular markers, which yielded clear and well-defined banding patterns. Furthermore, the extracted DNA was suitable for advanced applications, such as Sanger sequencing, by which high-quality electropherograms were obtained. The results demonstrate that the proposed protocol is an efficient, economical, and adaptable alternative for laboratories with limited resources, allowing the extraction of high-quality DNA from Gossypium barbadense L. and other plant species. This simplified approach facilitates the development of genetic and biotechnological research, contributing to the knowledge and valorization of the genetic resources of Peruvian native cotton.

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来源期刊
Methods and Protocols
Methods and Protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (miscellaneous)
CiteScore
3.60
自引率
0.00%
发文量
85
审稿时长
8 weeks
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