苎麻实时定量PCR体系的建立  

Establishment of Real-time Quantitative PCR System for Ramie

作　　者：马鑫[1] 谭松林[1] 邢虎成[1,2,3] 廖少波[1] MA Xin TAN Songlin XING Hucheng LIAO Shaobo(Ramie Institute of Hunan Agricultural University, Changsha 410128 , China Hunan Provincial Key Laboratory of Corp Germplasm Innovation and Utilization,Changsha 410128, China Grass Research Institute of Hunan Agricultural University, Changsha 410128, China)

机构地区：[1]湖南农业大学苎麻研究所,长沙410128 [2]湖南省种质资源创新与资源利用重点实验室,长沙410128 [3]湖南农业大学草业研究所,长沙410128

出　　处：《中国麻业科学》2017年第3期111-119,共9页Plant Fiber Sciences in China

基　　金：湖南省自然科学基金项目(2015JJ2083)

摘　　要：荧光定量PCR技术目前应用比较广泛,但是RNA质量、反转录效率、扩增效率和内参基因的选择等都会影响实时荧光定量PCR技术的准确性。研究以3个苎麻品种为材料,采用actin作为内参基因,利用2个苎麻基因进行实时定量PCR体系的建立。结果表明,采用试剂盒法提取的RNA可以满足RT-qPCR技术的要求,扩增曲线和熔解曲线都符合实验要求,actin基因可以作为内参进行苎麻相对定量分析研究。相对定量分析表明,研究建立的RT-qPCR方法可以应用于基因表达水平的研究。该研究为实时定量PCR在苎麻中的应用提供了参考。The application of real - time quantitative PCR technology is widely used at present. The accuracy of real - time quantitative PCR is affected by RNA quality and reverse transcription efficiency, amplification efficiency and reference gene. In this study, real - time quantitative PCR system was estab-lished using 3 ramie varieties, 2 ramie gene sequence, and actin gene sequence as the reference gene. The results showed that RNA extracted by RNAkits can meet the requirements of RT - qPCR. The ampli-fication curve and melting curve were consistent with the experimental requirements. Actin gene can be used as a reference for research on relative quantitative analysis of ramie. Relative quantitative analysis shows that the RT - qPCR method established in this study can be used to study gene expression level. This study can provide reference for the application of real - time quantitative PCR in ramie.

关 键 词：苎麻 实时定量PCR RNA 

分 类 号：S563.1[农业科学—作物学]