GSM8283558: scRNAseq_8cell_R48_7; Mus musculus; RNA-Seq
Znhit3+/- female mice 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 8-cell stage. When collecting samples, embryos were first washed in PBS and transferred into acidic Tyrode's solution to remove zonae pellucidae. Single zona-free embryo was then pipetted into a capillary tube with a diameter of 1.5 to 2 times of the diameter of single blastomeres to physically dissociate single blastomeres. Single blastomeres that remained intact were transferred into 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 scRNA-Seq library.