Gene regulation by CBP-p300 in the lens placode - E9.5

Wild type or CBP/p300 fx/fx; Le-Cre-positive (CBP/p300-/-) surface ectoderm from E9.5 mouse embryos was laser micodissected from three embryos of each genotype. Total RNA was purified, pooled for each genotype and triplicate samples were reverse transcribed and amplified using a NuGEN kit. cDNA was biotinylated and hybridized to Illumina Mouse6 v2.0 bead arrays. Three wild type and three knockout embryos were used. The RNA from embryos of the same genotype was purified and pooled and triplicate samples were amplified and used for microarray analysis.本实验针对两种基因型的E9.5期小鼠胚胎开展：野生型，以及CBP/p300 fx/fx; Le-Cre阳性（CBP/p300-/-）。从每类基因型的3枚胚胎中，通过激光显微切割（laser microdissection）分离得到表面外胚层组织。总RNA经纯化后按基因型分别混合，每份混合样品设置三个重复样本，使用NuGEN试剂盒完成反转录与扩增。将所得互补脱氧核糖核酸（complementary DNA，cDNA）进行生物素标记，随后与Illumina Mouse6 v2.0微珠芯片进行杂交。本次实验共纳入3枚野生型胚胎与3枚敲除型胚胎。同基因型胚胎的总RNA经纯化后混合，设置三个重复样本进行扩增并用于芯片分析。