快速高效提取荔枝幼果和离区组织RNA的新方法  

A New Rapid and Effective Method for RNA Isolation from Litchi Tissues of Fruitlet and Abscission Zone

作　　者：吴建阳[1] 彭刚[1] 李彩琴[1] 陆旺金[2] 王泽槐[1] 李建国[1] 

机构地区：[1]华南农业大学中国荔枝研究中心,广州510642 [2]华南农业大学园艺学院广东省果蔬保鲜重点实验室,广州510642

出　　处：《园艺学报》2011年第6期1191-1196,共6页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(30871694);教育部博士点基金项目(200805640003);国家现代荔枝产业技术体系项目(nycytx-32-03)

摘　　要：以荔枝幼果和离区为材料,以N-十二烷基肌氨酸钠(LSS)为RNA提取液的主要成分,建立了一种简单快速高效适合于荔枝幼果和离区总RNA提取的新方法,简称LSS法。该法提取RNA的过程只需2h,且质量高,幼果和果柄RNA的28S/18S值分别为1.7和1.9,RNA完整性(RIN)值分别为9.7和10.0,分别为380和450μg.g-1,A260/A280值分别为1.80和1.89。将提取的RNA经DNA酶消化后反转录成cDNA进行PCR,扩增出预期长度的目的片段,满足进行后续基因表达的要求。LSS法操作简单快速,对微量组织样品是一种有效的RNA提取方法。A new simple,rapid and effective LSS（N-lauroyl-sarcosine sodium）-based protocol for RNA isolation from litchi tissues of fruitlet and abscission zone was developed.This LSS method gave high-quality RNA in about 2 h.The ratios of 28S/18S,A260/A280,RNA integrity（RIN）and yield of the total RNA from tissues of fruitlet and abscission zone were 1.7 and 1.9,1.80 and 1.89,9.7 and 10.0,380 and 450 μg ？ g-1,respectivetly.In addition,reverse transcription-PCR confirmed that the isolated RNA was of appropriate quality and integrity for gene expression studies.Therefore,LSS method is particularly useful for processing large numbers of samples and for isolation of RNA fromminute quantities of tissue samples.

关 键 词：荔枝 幼果 离区 RNA提取 N-十二烷基肌氨酸钠(LSS) 

分 类 号：S667.1[农业科学—果树学]