Gene expression profiling of the tongue in mice with a conditional inactivation of Tgfbr2 at embryonic day E13.5

We performed global gene expression analyses of the tongue in Wnt1-Cre;Tgfbr2^fl/fl and Tgfbr2^fl/fl control mice at embryonic day E13.5. Total RNA samples (1 ug per sample) were converted into biotin-labeled cRNA using the Enzo BioArray Terminal Labeling Kit with Biotin-ddUTP and standard protocols recommended by Affymetrix (Santa Clara, CA). Fragmented cDNA was applied to GeneChip® Mouse Genome 430 2.0 Arrays (Affymetrix) that contain probe sets designed to detect over 39000 transcripts. Microarrays were hybridized, processed and scanned, as previously described using manufacturers recommended conditions.

本研究对胚胎期第13.5天（E13.5）的Wnt1-Cre;Tgfbr2^fl/fl条件性敲除小鼠与Tgfbr2^fl/fl对照小鼠的舌组织开展了全基因组基因表达分析。每份样本取1μg总RNA，采用搭载生物素-双脱氧尿苷三磷酸（Biotin-ddUTP）的Enzo BioArray终端标记试剂盒，并遵循Affymetrix（美国加利福尼亚州圣克拉拉市）推荐的标准实验流程，将总RNA转化为生物素标记的互补RNA（cRNA）。将片段化的互补DNA（cDNA）加载至GeneChip®小鼠基因组430 2.0芯片（Affymetrix），该芯片搭载的探针组可靶向检测超过39000种转录本。芯片杂交、样本处理及信号扫描均按照既往报道的方法，并严格遵循厂商推荐的实验条件完成。