title: GSM5048168: ChIP of SP1, replicate 2; Homo sapiens; ChIP-Seq
library_construction_protocol: ChIP (Chromatin Immunoprecipitation) was performed as previously described (Najafabadi et al., Nat. Biotechnol. (2015)). Briefly, ~20 million HEK293 cells were crosslinked in 1% formaldehyde. Lysates were sonicated to a DNA fragment length range of 200-300 bp using a Bioruptor (Diagenode). SP1 was immunoprecipitated from the lysate with 10ug of Sp1 antibody (Santa Cruz) and Dynabeads Protein G (Invitrogen). Subsequently, crosslinks were reversed at 65°C over night and bound DNA fragments were purified (EZ-10 Spin Column PCR Product Purification kit, Bio Basic). Sequencing libraries were constructed using the TruSeq ChIP-seq library kits (Illumina) according the manufacturer's instructions followed by PCR amplification and agarose gel size selection.