摘要
[目的]为了从团花树韧皮部中获取高质量的总RNA。[方法]研究采用冷酚法、一步提取法、改进CTAB法、杨树提取法、RNAplant Reagent法5种方法从团花树韧皮部组织中提取总RNA,并利用电泳拍照、测OD值和RT-PCR 3种方法对提取的RNA的质量进行检测。[结果]由冷酚法、一步提取法、改进CTAB法3种方法提取的RNA纯度较低,存在降解和弥散现象,或有DNA和蛋白质的污染,得到的RNA的量较少;而用杨树提取法和RNAplant Reagent法提取的RNA很少有降解,28SrRNA和18SrRNA条带很清晰,得到的RNA虽然有DNA的污染,但是经过DNaseⅠ处理后,OD260/OD280的值在1.8～2.0。[结论]杨树提取法和RNAplant Reagent法得到的RNA可以满足RT-PCR反应和RACE试验的要求,为成功克隆团花树相关基因奠定了基础。
[Objective] The research aimed to isolate high quality RNA from the phloem ofAnthocephalus chinensis. [Method] Several methods such as cold phenol method, one-step extraction method, improvement CTAB method, poplar extraction method and RNAplant Reagent method were used. The quality of total RNA was analyzed through gel electrophoresis, UV spectrometer, and RT-PCR.[Resuh] The result indicated that the purity of RNA isolated from the 3 methods of cold phenol method, one-step extraction method and improvement CTAB method were low, the RNA were degraded, dispersed in some degrees, or were polluted by DNA and protein, and less RNA were obtained. But the RNA obtained from the poplar extraction method and RNAplant Reagent method were rarely degenerated, and showed clear bands of 28S rRNA and 18S rRNA. Although the RNA were polluted by DNA, after dealing with the DNase Ⅰ, the value of OD260/OD280 reached between 1.8-2.0. [Conclusion] The RNA extracted from the poplar extraction method and RNAplant Reagent method could meet the request of RT -PCR reaction and RACE experiment, laying the foundation for the successful cloning genes of A nthocephalus chinensis.
出处
《安徽农业科学》
CAS
北大核心
2008年第11期4415-4418,共4页
Journal of Anhui Agricultural Sciences
基金
北京林业大学研究生自选课题基金(06jj058)
科学技术研究重点项目--高效集碳林木定向培育与能源化关键问题的基础研究(104243)
关键词
团花树
RNA提取
XET基因
Anthocephaluschinensis
RNA extractlon
XETgene
