GSM1572161: Ldb1_ChIP-seq; Mus musculus; ChIP-Seq
Cells were cross-linked with 1% formaldehyde in PBS at room temperature for 10 minutes. Chromatin was fragmented to 300-500 bp using a tip sonicator and centrifuged at 16,000 g for 10 min. The supernatant was pre-cleared with 20μl magnetic beads. The pre-cleared chromatin was incubated with 2-5 μg antibodies at 4°C overnight. Antibody-protein-DNA complexes were pulled down by incubating with 20μl Protein G Dynabeads (Life Technologies) and washed. The eluted complex was de-crosslinked overnight at 65°C. BLRP-tagged ChIP was performed using Nanolink Streptavidin Magnetic beads (Solulink) to directly pull down BLRP-tagged protein–DNA complex. Additional brief washes with 1% SDS in TE and TE buffer were introduced after normal washes. The washed streptavidin beads were then subjected to TEV protease (Life Technologies) digestion to release tagged protein and DNA complex before de-crosslinking at 65°C overnight. Extracted DNA was ligated to specific adaptors followed by deep sequencing on the Illumina GAII system according to the manufacturer’s instructions.
GEO
GSM1572161