不同保存方法对中麻黄基因组DNA提取效果的影响

摘要

The effect of genomic DNA extraction from Ephedra intermedia with different collection and preservation methods was compared and the best method was confirmed. The genomic DNA was extracted by improved CTAB method, the integrity, yield and purity of total genomic DNA were examined by UV-spectrophotomoter and agarose gel electrophoresis. High quality genomic DNA could be extracted from freshly frozen samples, whether the samples were frozen immediately or preserved in liquid nitrogen and silica gel after collection. The genomic DNA of fresh samples would degrade, which were frozen after drying a few days, and it was more serious that fresh samples were preserved in room temperature. The quality of genomic DNA would not be affected, which was brought back to laboratory by using different preservation methods, and cryopreservation could prevent genomic DNA degradation effectively.%通过对不同方法采集和保存的中麻黄样品基因组DNA提取效果进行比较,确定中麻黄的最佳采集和保存方法.应用改良CTAB法提取中麻黄基因组DNA,并采用琼脂糖凝胶电泳检测其完整性,利用紫外分光光度计法测定其浓度和纯度.结果表明,新鲜样品采集后立即冷冻保存或置于液氮和硅胶后冷冻保存,均能提取出高质量的基因组DNA；鲜样阴干几天后冷冻保存会导致基因组DNA降解；阴干后在室温下保存则降解更严重.采集的中麻黄鲜样采取不同方法带回实验室均不影响基因组DNA的提取效果,但应在低温环境中保存才能有效防止基因组DNA的降解.