建鲤IL–1β和IL–8基因实时荧光定量RT–PCR检测方法的建立.PDFVIP

() 2014 12 40 6 Journal of Hunan Agricultural University(Natural Sciences), Dec. 2014, 40(6):627–632 DOI:10.13331/ki.jhau.2014.06.013 /qks 建鲤 IL–1β和 IL–8 基因实时荧光定量 RT–PCR 检测方法的建立 1 1,2 1* (1. 625014 2. 614000) 摘 要GenBank β–actin SYBR Green I β–actinIL–1βIL–8 T 8682.5 84 ℃C 1232 m t 2 96.3%103.2%102.6%r 0.990 0.14%0.86%0.18% 0.93%0.13%0.86% IL–1βIL–8 1.09 1.71IL–1βIL–8 PCR IL–1βIL–8 关 键 词IL–1βIL–8 RT–PCR 中图分类号S941 文献标志码：A 文章编号：1007− 1032(2014)06−0627−06 Establishment of a real-time fluorescent quantitative RT–PCR for detection of IL–1β and IL–8 genes of Cyprinus carpio var. Jian 1 1,2 1* ZENG Dong , XIAO La , NI Xue-qin (1.Animal Microecology Institute, College of Veterinary, Sichuan Agricultural University, Ya’an, Sichuan 625014, China; 2.Animal Disease Prevention and Control Center of Leshan City, Leshan, Sichuan 614000, China) Abstract: To establish a real-time fluorescent quantitative PCR assay for detection of IL–1β and IL–8 genes of Cyprinus carpio var. Jian, specific primers were designed according to the gene sequences available in GenBank, and the β–actin gene was used as a reference gene. The standard curve and the melting curve were analyzed through SYBR Green I method. Melting curve showed specific PCR products of β–actin, IL–1β and IL–8 gene were obtained with Tm 86, 82.5 and 84 ℃ , respectively. Standard curve there were good linear relationship for β–actin, IL–1β and I