GSM3589850: NonInjection_S1: Non-injected 2-cell embryo -1; Mus musculus; RNA-Seq
A pool of five 2-cell embryos at 28 hpi after the removal of zona pellucida were subjected to RNA purification and cDNA synthesis using a SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (Takara Bio) according to the manufacturer's instructions. At the cell lysis step, 1 µL of ERCC (External RNA Controls Consortium) RNA Spike-In Control Mixes (1:40,000 dilution; Thermo Fishcer) was added to each of the samples. The cDNA was amplified with 11 cycles of PCR. For making a library for sequencing, Nextera XT DNA Library Preparation kit (illumina) was used and final PCR amplification was done for 13 cycles using standard Illumina protocols. The obtained library was quality-checked by Kapa Library Quantification kit (Nippon Genetics).