Tet-dependent transcriptional programs during early embryogenesis

To determine the transcriptional effects of lack of Tet proteins during early embryogenesis, we performed single-embryo RNA-sequencing of control and TKO embryos (E6.75; 4 embryos from each group). Genome-wide analyses showed that Tet deficiency promotes the expression of mesoderm-related genes during early embryogenesis in vivo. Control and Tet1/2/3 TKO embryos were collected at E6.75. Single embryos were lysed and converted into double stranded cDNA using SMARTer ultra-low RNA kit for Illumina sequencing (Clontech). 1 ng of cDNA for each sample was used for preparing libraries using Nextera XT DNA sample preparation kit (IIlumina). Libraries were sequenced on Illumina HiSeq 2500 platforms according to the manufacturer's instructions.