GSM9667520: PN0 mandibular molar tooth germs, Wnt1-Cre;Bmp2f/f mice, H3K4me3 (ChIP) 1; Mus musculus; ChIP-Seq
PN0 mandibular molar tooth germs were collected in ice-cold PBS, and each sequencing sample consisted of 10 tooth germs pooled as one biological replicate. Tissues were cross-linked with 1% formaldehyde for 10 min at room temperature and quenched with glycine (125 mM final concentration). Samples were lysed in buffer containing 50 mM Tris-HCl (pH 8.0), 10 mM EDTA, 1% SDS, and protease inhibitor cocktail, and chromatin was sheared by sonication to an average fragment size of 200-500 bp. ChIP-seq were performed by Wuhan IGENEBOOK Biotechnology Co.,Ltd (http://www.igenebook.com) .An aliquot of chromatin was reserved as DNA input, and immunoprecipitation was performed overnight at 4°C with rabbit anti-H3K4me3 (ABCam, ab8580; 10 μg antibody per reaction). Immune complexes were captured using protein beads and washed sequentially as described (low-salt, high-salt/LiCl, and TE washes). Bound chromatin was eluted in 100 mM NaHCO₃ with 1% SDS, treated with RNase A (final concentration 8 μg/mL) at 65°C, followed by proteinase K digestion (final concentration 345 μg/mL) at 45°C. ChIP DNA was purified and used to prepare sequencing libraries using the NEXTFLEX® ChIP-Seq Library Prep Kit for Illumina® Sequencing (NOVA-5143-02, Bioo Scientific).