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当前位置: 首页 > 文章 > 油菜蜂花粉总RNA(含小RNA)大量提取及RT-qPCR检测 中国蜂业 2016,67 (10) 18-20
Position: Home > Articles > Mass extraction of total RNA(containing small RNA) and RT-qPCR detection of rape(Brassica napus) bee pollen Apiculture of China 2016,67 (10) 18-20

油菜蜂花粉总RNA(含小RNA)大量提取及RT-qPCR检测

作  者：
陈璇;吴人照;王刚;龙华晴;戴关海;任泽明;童晔玲;杨锋
单  位：
浙江省中医药研究院基础实验研究所;浙江省中医药研究院中西医结合肿瘤研究所
关键词：
油菜蜂花粉;总RNA大量提取;miRNA;RT-qPCR
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。
译  名：
Mass extraction of total RNA(containing small RNA) and RT-qPCR detection of rape(Brassica napus) bee pollen
作  者：
Chen Xuan;Wu Renzhao;Wang Gang;Long Huaqing;Dai Guanhai;Ren Zeming;Tong Yeling;Yang feng;Institute of Basic Medicine, Zhejiang Academy of Traditional Chinese Medicine;Institute of Traditional Chinese Medicine and Western Medicine in Cancer Research,Zhejiang Academy of Traditional Chinese Medicine;
关键词：
rape bee pollen;;mass extraction of total RNA;;miRNA;;RT-qPCR
摘  要：
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.

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