GSM5352789: 8Cell H3K9me3 Input; Homo sapiens; ChIP-Seq
To obtain MII oocytes, the immature oocytes were collected and matured in vitro in medium (Cooper Surgical/SAGE, Trumbull, CT) supplemented with 75 mIU/ml FSH and LH for 24-28h. After the in vitro maturation, the MII oocytes were fertilized by intracytoplasmic sperm injection (ICSI), and then cultured in IVF medium (Vitrolife, catalog. no. 10136). 16-18h later, the fertilization were observed, 2PN zygotes were cultured in G1 medium (Vitrolife) at 37°C in 6% CO2, 5% O2 and 89% N2 for 3 days to 8-cell embryo stage, and then switched to G2 medium (Vitrolife) until the blastocyst stage. The 4-cell, 8-cell and morula embryos (zona pellucida and Polar body removed) were collected into the nuclear isolation buffer for AULIChIP-seq. The inner cell mass (ICM) and mural trophectoderm were separated by gently pipetting in a pipette with a diameter of 40-60 μm and collected into the nuclear isolation buffer.  Human ESCs were dissociated with Accutase (Thermo Fisher Scientific, catalog. no. A1110501) into single cells and washed with PBS solution twice. Then, 50 or 500 hESCs were collected into the nuclear isolation buffer.   Lysates were clarified from micrococcal nuclease-treated nuclei and histone-DNA complexes were isolated with anti-H3K9me3 antibody. Libraries were prepared according to the Accel-NGS  2S Plus DNA Liibrary Kit from Swift Biosciences.