GSM7072062: BDF1_in-vitro_G1_04-cell_DEP-01-029-23; Mus musculus; OTHER
Single-cell isolation: Collection of single blastomeres was performed using micromanipulation. Embryos were transferred into M2 medium supplemented with 5 μg/ml cytochalasin B (Sigma-Aldrich) for 10 min. Then, the zona pellucida was cut by the LYKOS laser system (Hamilton Thorne) in a micromanipulation chamber, which was put on a warmed stage (37°C) in an inverted microscope (Olympus Co.). After cutting the zona pellucida, a fire-polished injection pipette (inner dia. 30 µm) was inserted through the hole of the zona pellucida, single blastomere was isolated. For late 8-cell and 16-cell stage embryos, embryos were treated with TrypLE Express (gibco) for 5 min before single-cell isolation. TE and ICM cells were isolated described previously (Xu et al., 2013) and microsurgical isolation to single cell was performed by micromanipulator.  After the single-cell isolation, sample preparations were performed as described (Miura et al., 2020). DNA samples were subject to whole-genome amplification (WGA) with a SeqPlex kit (Sigma, SEQXE). NGS libraries were constructed WGA DNA with an NGS LTP Library Preparation Kit (KAPA, KK8232) according to the manufacturer's instructions