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DOI: 10.3791/1169-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
This protocol outlines the process for vaccinia virus infection of HeLa cells and subsequent analysis of both host and viral gene expression. The focus is on synthesizing cDNA from isolated RNA for further analysis.
协议牛痘感染的HeLa细胞和宿主和病毒基因的表达分析。
大家好,我是来自怀特黑德生物医学研究所 Kate Rubins 实验室的 Judy Yen。用牛痘病毒感染 hela 细胞并分离总 RNA 后,如之前的 Joe 视频方案帖子所示,可以使用反向和体外转录分离和扩增病毒 RNA,扩增的 RNA 稍后将用于病毒和宿主基因表达的微阵列分析。那么让我们开始吧。
为了开始从 RNA 合成 c cd NA,在洗涤缓冲液中使用环境氨基等位基因信息 AMP two 试剂盒之前,将推荐体积的 100% 乙醇加入洗涤缓冲液中,在 PCR 反应管中加入 100 ng 至 5 μg 总 RNA 和 1 μL T 7 寡核苷酸引物。用不含核酸酶的水将体积增加到 12 μL。将样品在 70 摄氏度下在热循环仪中孵育 10 分钟。
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