改良热酚法快速提取棉花不同组织RNA

Rapid Extraction of Total RNA from Different Cotton Tissues by Modified Hot-phenol Method

[目的]以改良热酚法快速提取棉花不同组织的RNA。[方法]以陆地棉(Gossypium hirsutum L.)新陆早26为材料,在冷酚法的基础上对RNA的提取方法进行改进。[结果]利用改良热酚法获得的RNA不仅完整性极好、浓度高且无多糖多酚污染,可用于cDNA末端快速扩增(RACE)、Northern Bloting、基因芯片分析及转录组分析等研究。[结论]该方法不但适用于提取棉花叶片总RNA,而且在根、花冠、花药、发育的纤维中均能获得高质量的RNA,同时节约成本,操作简便,对环境污染小。

[Objective] This study aimed to develop a hot-phenol method for extracting the total RNA from different cotton tissues.[Method] The total RNA of different tissues of Gossypium hirsutum cultivar Xinluzao 26 was extracted with cold-phenol method,Trizol method,modified hot-phenol method and CTAB method,respectively;all the extracted RNA samples were analyzed with electrophoresis and compared.[Result] The RNA samples extracted by modified hot-phenol method were complete,high-concentration,free from contaminations by polysaccharides and polyphenols,and could be used for rapid-amplification of cDNA ends,northern blot,gene chip analysis and transcriptome analysis,etc.[Conclusion] This method can be adopted to extract high-quality total RNA from cotton leaves,root,shoot,corolla,anther,ovule and developing fibers.Meanwhile,it is simple,low-cost and produces little pollution to environment.