GSM7371843: LacZ - control - Input; Homo sapiens; ChIP-Seq
Cells were cross-linked with 1% formaldehyde for 10 minutes at RT, followed by quenching with glycine (final concentration 125 mM). After washing with ice-cold PBS, cells were collected through centrifugation (400g). For the analysis of total lamin A/C chromatin binding, cells were lysed and chromatin was prepared using the Chromatin EasyShear Kit - High SDS (Diagenode; Cat# C01020012), and sonicated using an E220 focused ultrasonicator (Covaris). The ChIP reaction was performed using the iDeal ChIP-seq kit (Diagenode, Cat# C01010055). Samples were pre-cleared using the beads provided in the kit and incubated overnight at 4°C with 5ug of anti-Androgen Receptor, PG-21, Cat. No. 06-680 (Sigma). Antibody-chromatin complexes were pulled down using protein A-coated magnetic beads. Elution was performed as per the manufacturer's instructions and chromatin was quantified using the Qubit Fluorometric Quantification Kit (Thermo Scientific). For library preparation, 10 ng of DNA was utilized with the NEBNext® ChIP-Seq Library Prep Reagent Set for Illumina. Subsequently, sequencing was performed at the Novegene facility using the Novaseq 6000 platform. For library preparation, 10 ng of DNA was utilized with the NEBNext® ChIP-Seq Library Prep Reagent Set for Illumina.