Whole-genome transcriptome profiling of E13.5 Wnt1-Cre;Alk5flfl by microarray of maxilla and mandible

We performed the global gene expression profiling analysis of the proximal (P ) and distal (D) regions of the mandible and proximal (P ) and distal (D) regions of the maxilla using microarray. Total RNA samples (1 ug per sample) were converted into biotin-labeled cRNA using the GeneChip® IVT Labeling Kit and standard protocols recommended by Affymetrix (Santa Clara, CA). Fragmented cRNA was applied to GeneChip, Mouse Genome 430 2.0 Arrays (Affymetrix) that contain probe sets designed to detect over 39,000 transcripts. Microarrays were hybridized, processed and scanned, as previously described using manufacturers recommended conditions.

本研究采用微阵列（microarray）技术，对下颌骨（mandible）的近端（P）与远端（D）区域，以及上颌骨（maxilla）的近端（P）与远端（D）区域开展全基因表达谱分析。每份样品取1 μg总RNA，使用GeneChip®体外转录标记试剂盒（GeneChip® IVT Labeling Kit）及Affymetrix公司（加利福尼亚州圣克拉拉市）推荐的标准实验流程，将总RNA制备为生物素标记的互补RNA（cRNA）。将片段化后的cRNA加载至小鼠基因组430 2.0芯片（Mouse Genome 430 2.0 Arrays，Affymetrix），该芯片搭载有可检测超过39000种转录本的探针集。微阵列实验的杂交、处理与扫描流程均参照既往研究方案及厂商推荐的实验条件完成。