一种快速提取猕猴桃RNA的新方法  

A New Method for Rapid Extraction of RNA in Kiwi

作　　者：邓浪 包昌艳[1,2] 周军 王连春[1,2] 刘惠民 纪晴[1,2] 沈兵琪 周凡[1,2] 王大玮 Deng Lang;Bao Changyan;Zhou Jun;Wang Lianchun;Liu Huimin;Ji Qing;Shen Bingqi;hou Fan;Wang Dawei(Key Laboratory of Biodiversity Conservation of the State Forestry Administration of Southwest China, Kunming, 650244;Key Laboratory for Forest Resources Conservation and Use in the Southwest Mountains of Education, Southwest Forestry University, Kunming, 650244;College of Biologieai Sciences and Engineering, North Minzu University, Ningxia, 750021)

机构地区：[1]西南地区生物多样性保育国家林业局重点实验室,昆明650224 [2]西南林业大学,西南山地森林资源保育与利用省部共建教育部重点实验室,昆明650224 [3]北方民族大学生物科学与工程学院,宁夏750021

出　　处：《分子植物育种》2018年第10期3234-3239,共6页Molecular Plant Breeding

基　　金：国家林业局948项目(2012-4-62)资助

摘　　要：本研究通过参考CTAB法、异硫氰酸胍法、Trizol法、SDS法以及"快速从含有高含量淀粉的种子胚乳中分离高质量RNA"方法,以‘Hort-16A’的幼叶、花和幼果进行总RNA提取,并对上述方法的结果进行比较。在综合上述方法进行改进后,筛选出了快速、稳定、操作简单、成本低,适合猕猴桃总RNA提取方法。与所参考的几种方法比较,该新方法提取RNA完整清晰。将提取RNA反转录c DNA为模板,以AP2基因和AG基因设计两对特异引物进行RT-PCR扩增,获得条带明亮单一,完整,经测序为所需目的条带。该新方法提取的RNA可为下一步进行基因克隆和功能验证实验提供数据资料。In this study, the total RNA was extracted from young leaves, flowers and young fruits of‘Hort-16 A＇by reference to CTAB method, guanidine thiocyanate method, trizol method, SDS method and＂rapid separation of high quality RNA from seed endosperm containing high levels of starch＂method, and the results of the reference methods were compared. After the improvement of the above methods, the method of rapid, stable, simple operation and low cost was selected which was suitable for the total RNA extraction of kiwifruit. Compared with several methods referenced, RNA extracted in the new method was integrated and clarified. Using the reverse transcription c DNA of extracted RNA as template, two pairs of specific primers designed from AP2 gene and AG gene were applied for RT-PCR amplification. The stripe obtained was bright, single and complete, which was the required purpose strip after sequencing. The RNA extracted by this new method might provide data information for the next step in gene cloning and functional verification experiments.

关 键 词：Hort-16A 猕猴桃 RNA RT-PCR 

分 类 号：S663.4[农业科学—果树学]