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.%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ac-tin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.
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