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《China Animal Husbandry & Veterinary Medicine》 2014-03
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Construction of Small-tailed Han Sheep Leptin Receptor Gene Standard Plasmid and Standard Curve Using Real-time Quantitative PCR

ZHANG Gui-shan;XU Jing;LOU Yu-jie;ZHANG Shan-peng;JIANG Huai-zhi;College of Animal Science and Technology,Jilin Agricultural University;Changchun University of Science and Technology;  
In order to prepare the leptin receptor long form gene standard plasmid and standard curve using Real-time quantitative PCR,the total RNA was isolated from the arcuate nucleus tissue of ovis aries before puberty,and was synthesized the first-strand cDNA via reverse transcription.Using PCR technology,the leptin receptor long form gene was amplified.The aim gene fragment was recycled,and the fragment was then cloned into the pMD18-T plasmid vector.Recombinant plasmid of recycled products was constructed.After extraction and identification,the recombinant plasmids were quantified and the standard curve was established.The amplified products of the recombinant plasmids and secquence analysis confirmed that the pMD18-Tleptin receptor long form gene was successfully cloned.The standard curve showed high linearity,sensitivity,specificity and exercisable,which can be used to detect leptin receptor long form gene expression.
【Fund】: 吉林省科技支撑计划项目(20110237)
【CateGory Index】: S826
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