RNASeq analysis of embryonic day 13.5 mouse cerebral cortex from (i) Pax6-deleted and (ii) Pax6- and Gsx2-deleted embryos

Mice carrying embryos that were either Pax6fl/fl Gsx2fl/+ Emx1CreER or Pax6fl/fl Gsx2fl/fl Emx1CreER were given tamoxifen on embryonic day 9.5 (E9.5) to induce either Pax6 deletion alone or Pax6 and Gsx2 deletion together. Embryos were harvested on E13.5, the cerebral cortices were removed and divided into rostral and caudal halves, and total RNA was extracted from the rostral half. Samples from littermates of the same genotype were pooled. Poly-A mRNA was purified and TruSeq RNA-Seq libraries were prepared and sequenced (100 base paired-end; Illumina, HiSeq v3).

携带两种指定基因型胚胎的小鼠，其基因型分别为Pax6fl/fl Gsx2fl/+ Emx1CreER与Pax6fl/fl Gsx2fl/fl Emx1CreER，于胚胎第9.5天（E9.5）予以他莫昔芬处理，分别诱导单独Pax6基因敲除，或同时敲除Pax6与Gsx2基因。随后在胚胎第13.5天（E13.5）收获胚胎，剥离其大脑皮层并分为吻侧与尾侧两半，取吻侧半段提取总RNA。将同基因型同窝胚胎的样本混合后，纯化得到聚腺苷酸mRNA（Poly-A mRNA），构建TruSeq RNA测序文库并进行测序（100碱基双端测序；采用Illumina HiSeq v3平台）。