Lopez Luis Miguel Serquén, Gonzales Herry Lloclla, Garcia Wilmer Enrique Vidaurre, de Jesus Velez Chicoma Ricardo Leonidas, Greta Mendoza Cornejo
Research Center, Professional School of Environmental Engineering, César Vallejo University, Lambayeque 13001, Peru.
Methods Protoc. 2025 May 7;8(3):50. doi: 10.3390/mps8030050.
Efficient extraction of high-quality DNA from plants is a critical challenge in molecular research, especially in species such as 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 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 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.
从植物中高效提取高质量DNA是分子研究中的一项关键挑战，尤其是对于原产于秘鲁的[植物名称]等物种而言，因为其存在多糖和酚类化合物等抑制剂。本研究提出了一种基于CTAB并结合硅胶柱的改良方案，旨在克服这些限制，且无需液氮或昂贵试剂。在秘鲁兰巴耶克采集了本地棉花样本，并采用简化程序进行处理，该程序优化了提取DNA的纯度和浓度。对八个具有彩色纤维（米色、fifo、浅棕色、深棕色、橙棕色、微红、细微红和白色）的[植物名称]品种进行了评估，得到的DNA的A260/A280比值在2.14至2.19之间，A260/A230比值在1.8至3.14之间；这些值高于用经典CTAB方法获得的值。通过使用ISSR和RAPD分子标记的PCR扩增验证了DNA质量，得到了清晰且明确的条带模式。此外，提取的DNA适用于诸如桑格测序等高级应用，通过该应用获得了高质量的电泳图。结果表明，所提出的方案对于资源有限的实验室而言是一种高效、经济且适应性强的替代方案，能够从[植物名称]和其他植物物种中提取高质量DNA。这种简化方法促进了遗传和生物技术研究的发展，有助于对秘鲁本地棉花的遗传资源进行了解和增值。
