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肝组织中β-actin定量检测方法的建立 被引量:1

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摘要 目的构建检测肝组织中内参基因β-actin的标准品。方法以成人外周血细胞的总RNA为模板、反转录合成cDNA,用该cDNA为模板PCR扩增人β-actin基因相应的cDNA目的片段,以pMD18-T为载体,构建形成重组质粒,电泳、鉴定测序后,用荧光定量PCR制作标准曲线。结果外周血提取的总RNA完整性良好,构建的β-actin质粒经PCR扩增后得到1条285 bp的清晰条带,测序结果与目的片段完全一致,且质粒的原始浓度为5.0×10^(13)拷贝数/mL,倍比稀释至1.0×10~2拷贝数/mL均能得到良好的标准曲线(R^2=0.999)。结论构建β-actin基因荧光定量PCR标准质粒成功。
出处 《肝脏》 2017年第7期629-631,共3页 Chinese Hepatology
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