Gene microarray screening of RNA from bruce 4 ES cells. Mus musculus

The sample is wild type male bruce 4 ES cells which has been used in core f to generate knockout mice. Overall design: RNA samples from wild type male bruce 4 ES cells, which has been used in core f to generate knockout mice, were screened. Scraped cells, pre-plated and non-pre-plated trypsinized cells were compared to determine the effects of trypsin and fibroblasts on gene expression of the ES cells. Mouse ES cell pellets were delivered to Core E for RNA Extraction. *Concurrent analysis of glycan profiles in these cells was performed in Core C*. RNA from scraped ES cells, trypsinized cells, and pre-plated trypsinized cells was isolated and prepared in triplicate. RNA was labeled and hybridized to the GLYCOv3 array. Resulting Gene expression patterns were analyzed by Core F.

本数据集的样本为野生型雄性Bruce 4 ES细胞（Embryonic Stem Cells），该细胞曾被用于核心实验室F构建基因敲除小鼠。
实验设计概述：本研究对曾用于核心实验室F构建基因敲除小鼠的野生型雄性Bruce 4 ES细胞的RNA样本进行了筛选。通过对比刮取细胞、预铺板胰酶消化细胞与未预铺板胰酶消化细胞，以明确胰酶与成纤维细胞对ES细胞基因表达的影响。小鼠ES细胞沉淀被送至核心实验室E进行RNA提取。*本研究同时在核心实验室C完成了这些细胞的糖基化谱分析*。对刮取的ES细胞、胰酶消化细胞及预铺板胰酶消化细胞的RNA进行了三次重复的分离与制备。RNA经标记后与GLYCOv3基因芯片（GLYCOv3 array）进行杂交。最终得到的基因表达谱由核心实验室F完成分析。