RNA Seq analysis of e12.5 mouse pancreatic buds from control and Pdxcre; Gata4fl/fl;Gata6fl/fl; Tom mice

about 250 genes were significantly changed after Gata4 and Gata6 were specifically deleted in the pancreatic progenitor cells 6 pancreatic buds were pooled for the control, and 12 pancreatic buds were pooled for the Pdxcre; Gata4fl/fl; Gata6fl/fl. Libraries were prepared from total RNA (RIN>8) with the TruSeq RNA prep kit (Illumina) and sequenced using the HiSeq2000 (Illumina) instrument. More than 20 million reads were mapped to the mouse genome (UCSC/mm9) using Tophat (version 2.0.4) with 4 mismatches and 10 maximum multiple hits. Significantly differentially expressed genes were calculated using DEseq在胰腺祖细胞中特异性敲除Gata4与Gata6基因后，约有250个基因发生显著表达变化。对照组合并6个胰腺芽，Pdxcre; Gata4fl/fl; Gata6fl/fl实验组则合并12个胰腺芽。以总RNA（RNA完整性指数RIN>8）为原料，使用Illumina公司的TruSeq RNA建库试剂盒构建测序文库，并通过Illumina HiSeq2000测序仪完成测序。采用Tophat软件（版本2.0.4），允许4个碱基错配、最多10个多重比对位点，将超过2000万条测序读段比对至小鼠参考基因组（UCSC/mm9）。通过DEseq软件计算得到显著差异表达基因。