南瓜DNA提取方法比较分析

摘要

DNA提取是基因克隆、PCR扩增、分子杂交以及遗传多态性分析等实验的基础.Whatman FTA卡已广泛应用于DNA的保存和提取,具有简便和高效的特点.采用煮沸法和FTA法分别提取南瓜(Cucurbita moschata Duch.)根、茎、叶DNA,以常规CTAB法为对照,以期寻找一种简便快捷的DNA提取方法.结果表明,CTAB法适用于南瓜各组织部位的DNA提取,DNA质量最高,PCR扩增的条带清晰,亮度高,但实验操作也最复杂;用煮沸法从叶片中提取的DNA质量相对较好,从根和茎提取的DNA不宜用于PCR反应,实验操作相对简单;FTA法提取的DNA质量较好,PCR扩增的条带清晰,亮度较高,实验操作最简单,是一种最佳的DNA简化提取方法.%The DNA extraction is a basic experimental protocol of gene cloning,PCR reaction,molecular hybridization and genetic polymorphism analysis.Whatman FAT card has been widely applied on DNA conservation and extraction with high efficiency and simplicity.In this study,pumpkin (Cucurbita moschata Duch.) root,stem and leaf were used to extract DNA using boiling method and FTA method,using CTAB method as reference,aiming to explore a simple and efficient DNA extraction method.The results indicated that the DNA quality extracted by using CTAB method was optimal compared to the other two methods and the bands were clear and bright.However,CTAB method was the most complicated.The DNA quality extracted from boiling method,except leaf,was poor and not fit for PCR even though the protocols was relatively simple.The DNA quality extracted using FAT method was good and fit for PCR with clear and bright bands.FTA method is optimal since the protocol is simple and efficient.