GSM8675922: 2cell_P2_110723_cDNA_C12_S98; Mus musculus; RNA-Seq
Mouse zygotes at E0.5 were extracted from super ovulated C57/Bl6 mice. Zygotes were cultured in vitro until 2-cell and 4-cell stages. Zygotes/1-cell embryos were directly transferred into RLT buffer in 96-well plates with 1 zygote/ well. Single cells from 2-cell embryos were extracted 26-28 hours after zygote extraction. Single cells from 4-cell embryos were extracted 38-40 hours after zygote extraction. For 2-cell and 4-cell stages, first the zona pellucida was removed using the Tyrode Acid Solution, then the embryos were incubated in TrypLE: Accutase (1:3) solution for ~10 minutes until the embryos dissociated into single cells. Single cells were then mouth pipetted into individual wells of a 96-well plate containing RLT buffer. The wells were spun down and stored at -20C until further processing. Nextera XT DNA Library Prep Kit was used for for each sample. Nextera XT 96-index kit was used to barcode each sample. All samples were pooled together before sequencing on the same chip.