Echevarría-Machado Ileana, Sánchez-Cach Lucila A, Hernández-Zepeda Cecilia, Rivera-Madrid Renata, Moreno-Valenzuela Oscar A
Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán, A. C. Calle 43 No. 130, Col. Chuburná de Hidalgo, CP 97200, Mérida, Yucatán, México.
Mol Biotechnol. 2005 Oct;31(2):129-35. doi: 10.1385/MB:31:2:129.
A protocol is described for rapid DNA isolation from Malvaceae plant species and different tissues of Bixaceae that contain large amounts of polysaccharides, polyphenols, and pigments that interfere with DNA extractions. The method is a modification of Dellaporta et al. The current protocol is simple, and no phenol-chloroform extraction, ethanol, or isopropranol precipitation is required. The method is based in the incubation of soluble DNA with silica, mix in batch during the extraction. The procedure can be completed in 2 h and many samples can be processed at the same time. DNA of excellent quality was recovered and used for polymerase chain reaction (PCR) amplification, restriction enzyme digestion, and Southern blot analysis. The method was used with healthy Bixa orellana and virus-infected Malvaceae plants.
本文描述了一种从锦葵科植物物种以及含有大量干扰DNA提取的多糖、多酚和色素的红木科不同组织中快速分离DNA的方法。该方法是对Dellaporta等人方法的改进。当前方案简单，无需酚 - 氯仿萃取、乙醇或异丙醇沉淀。该方法基于可溶性DNA与硅胶孵育，在提取过程中分批混合。该过程可在2小时内完成，并且可以同时处理多个样品。回收了高质量的DNA并用于聚合酶链反应（PCR）扩增、限制性内切酶消化和Southern印迹分析。该方法应用于健康的红木和感染病毒的锦葵科植物。
