title: GSM8762523: 4C_ZF-LDB1-DD_in_SSBP3-dTAG_DMSO_rep3; Mus musculus; OTHER
library_construction_protocol: 10 million cells were fixed. DpnII and NlaIII were used as first and second restriction enzyme, respectively. Overnight ligation was performed after digestion. The following 4C primers were used to amplify viewpoints of interest, using 200 ng 4C template input in 4 independent PCR reactions (4 x 50 μl): β-major promoter TACACGACGCTCTTCCGATCTGCAGAGCATATAAGGTGAGGTAGGATC and ACTGGAGTTCAGACGTGTGCTCTTCCGATCTCTTTTCTTTGTTTCTCAGTTTGAGTGCATG Pooled reactions were purified by QIAquick PCR Purification Kit (QIAGEN, 21804) eluting in 50 μl milliQ water. 10 μl purified PCR product was used as input material for a second PCR to obtain full Illumina adapter sequences using a universal forward primer (AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT) and a barcoded reverse primer (CAAGCAGAAGACGGCATACGAGATXXXXXXGTGACTGGAGTTCAGACGTGTGCT) with a 6-nucleotide barcode (XXXXXX). Pooled 4C libraries were purified with Ampure XP beads (Beckman Coulter, A63881) before sequencing on the Illumina NextSeq 2000 platform.