Instrument: Illumina HiSeq 2000
Strategy: OTHER
Source: GENOMIC
Selection: other
Layout: SINGLE
Construction protocol: The initial steps of the 4C procedure, as published before (Simonis et al., 2006, Nature Genetics), remain unchanged. Cells are cross linked using 2% formaldehyde for 10min at room temperature in PBS 10%FCS, nuclei are isolated, after which chromatin is digested with DpnII or NlaIII and subsequently ligated. After reversal of the cross links the DNA is purified and treated with the second restriction enzyme treatment (Csp6I or NlaIII, respectively). After a second re-ligation step the sample is purified and a 4C PCR is performed.