柿属植物叶绿体 DNA 提取方法优化

摘要

利用 Percoll 密度梯度离心法，对4种柿属植物君迁子 D. lotus L.、油柿 D. oleifera Cheng、浙江柿 D. glaucifolia Metc.、金枣柿 D. kaki spp.进行叶绿体 DNA（cpDNA）提取方法的优化，结果显示优化后的柿属植物cpDNA 提取方法具有高效、简便、重复性好、稳定性高的特点；经检验，利用此方法提取的 cpDNA 质量好，浓度与纯度均较高，能够满足后续叶绿体 PCR 扩增、基因组测序等分子生物学试验。%In order to optimize the method of extracting chloroplast DNA(cpDNA)from Diospyros lotus L., D. oleifera Cheng, D. glaucifolia Metc. and D. kaki spp., the technology of Percoll density-gradient centrifugation was used. The results show that the cpDNA of Diospyros could be quickly, simply, repeatedly and steadily extracted by the method. The quality of the cpDNA which was isolated by this method was determined through the technology of agarose gel electrophoresis, which indicated that the purity, concentration and quality of cpDNA was good enough for further PCR amplification and genomic sequencing analysis.