油菜蜂花粉总RNA(含小RNA)大量提取及RT-qPCR检测  

Mass extraction of total RNA(containing small RNA) and RT-qPCR detection of rape(Brassica napus) bee pollen

作　　者：陈璇[1] 吴人照[1] 王刚[2] 龙华晴 戴关海[1] 任泽明[1] 童晔玲[1] 杨锋[1] 

机构地区：[1]浙江省中医药研究院基础实验研究所,310007 [2]浙江省中医药研究院中西医结合肿瘤研究所,310007

出　　处：《中国蜂业》2016年第10期18-20,共3页Apiculture of China

基　　金：浙江省自然科学基金青年基金(LQ13C170002);浙江省科技计划项目科技条件建设(2013F10001)

摘　　要：大量提取含小RNA的油菜蜂花粉总RNA,为研究油菜蜂花粉所含外源性miRNA在动物和人体内发挥的生理和病理功能研究奠定基础。改良苯酚-氯仿法大量提取油菜蜂花粉总RNA,Nanodrop 2000检测浓度和质量,琼脂糖凝胶电泳检测RNA完整性;所得的总RNA经过RT-qPCR扩增。提取的总RNA260/280为2.11,产量高于1‰。电泳条带清晰,完整性良好。可扩增出油菜基因组中的miR-159和miR-166a,且可进行丰度差异比较。本方法能大量提取包括油菜蜂花粉在内的植物RNA(含小RNA),为中草药和其他植物来源天然产物中功能性miRNA在动物和人体内发挥的生物学功能研究奠定基础。Total RNA that contain miRNAs were extracted from rape bee pollen, which laid a foundation to study the physiological and pathological functions of exogenous miRNAs of rape bee pollen in animals and human. Total RNA from mass rape bee pollen were extracted using a modified phenol-chloroform method, and the concentration and quality were determined by Nanodrop2000 as well as electrophoresis with 1% agarose gel. The total RNA was amplified by RT-qPCR. The ratios of A260/A280 of total RNA was 2.11, and the yield was over 1‰. The bands were clear on agarose gel and the integrity of RNA was good. Different abundance of miR-159 and miR-166a were amplified. The method can extract RNA （containing small RNA） from mass plant including rape bee pollen, which laid a foundation to study biological functions of functional miRNAs from herb and other natural products of plant origin in animals and human.

关 键 词：油菜蜂花粉 总RNA大量提取 MIRNA RT—qPCR 

分 类 号：S896.4[农业科学—特种经济动物饲养]