MSCs and meshes for Hernia - qPCR data.txt

Gene expression of cells infiltrated in the surgical meshes at 1 week and 1 month after surgical mesh implantation was analyzed by quantitative Real Time PCR (qPCR). qPCR was performed with commercial gene expression assays (Life Technologies, Thermo Fisher Scientific Inc.), using TaqMan Fast Advance Master Mix (Life Technologies, Thermo Fisher Scientific Inc.), in accordance with the protocol provided by the manufacturer. The amplification of cDNAs was performed in a QuantStudio 3 System (Applied Biosystems, Thermo Fisher Scientific) and the qPCR products were quantified by fluorescent method using the 2^-ΔCt expression. Duplicates of all samples were analyzed separately and normalized using HRPT1 as a reference gene.

本研究采用实时定量聚合酶链式反应（quantitative Real Time PCR，qPCR），对外科植入补片植入后1周及1个月时，浸润至补片内的细胞的基因表达水平进行分析。实验使用赛默飞世尔科技公司（Thermo Fisher Scientific Inc.）旗下Life Technologies品牌的商业化基因表达检测试剂盒，搭配TaqMan Fast Advance预混液（TaqMan Fast Advance Master Mix），严格遵循厂商提供的实验方案完成qPCR反应。互补脱氧核糖核酸（complementary DNA，cDNA）的扩增反应在QuantStudio 3型荧光定量PCR系统（Applied Biosystems，赛默飞世尔科技公司）中完成，qPCR产物通过荧光法定量，采用2^-ΔCt法计算基因表达量。所有样本均设置技术重复并单独分析，以HRPT1作为内参基因完成数据归一化处理。