GSM2068817: Lamin B1 young replicate 2; Mus musculus; ChIP-Seq
Snap-frozen mouse liver (100 mg) from wild type mice was used to prepare chromatin. Foxa2-specific rabbit antiserum (Seven Hills Bioreagents, WRAB-1200), rabbit antibody to Di Methyl-Histone H3 Lys9 (Cell Signaling, D85B4), goat antibody to lamin B1 (Santa Cruz Biotechnology, sc-6216) and rabbit anti-histone H3 (trimethyl K9) antibody (ab8898) were used for immunoprecipitation. Libraries were prepared according to Illumina's instructions. Briefly, DNAs were blunted with a combination of T4 DNA polymerase, Klenow polymerase, and T4 PNK, then a single 3′-end “A” base was added using Klenow exo (3′-to-5′ exo minus). Broad custom adapters were then ligated to the ends of the modified DNA.  After adapter ligation DNA was PCR amplified with Broad custom primers for 16 cycles and library fragments of ~280 bp (~150-bp  insert plus ~130-bp adaptor and PCR primer sequences) were  isolated using Pippin Prep agarose gel. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on Illumina Hiseq 2500 following the manufacturer's protocols.