GSM8283628: Znhit3_4cell_se22; Mus musculus; RNA-Seq
To collect embryos, Znhit3+/- females were injected intraperitoneally with eCG (5 IU) followed 46 h to 48 h with hCG (5 IU) and then co-caged with Znhit3+/- males. Fertilized zygotes were flushed from plugged females 16 h post hCG injection and cultured in M2 medium containing hyaluronidase to remove the cumulus mass. Zygotes without cumulus were then washed and cultured in advanced KSOM until sample collection. When collecting samples, embryos were washed in PBS and transferred into acidic Tyrode's solution to remove zonae pellucidae. Single zona-free embryos were transferred into 8-well PCR strips containing 2.5 µl of RLT plus buffer. RNAs in each sample were captured by oligo-dT beads and then processed following the G&T-seq protocol to construct the single embryo RNA-Seq library.