GSM6181490: 4C_St3gal6.TSS_Dox_Untreated_rep1 [722]; Mus musculus; OTHER
4C-seq experiments were performed as previously described (Krijger PHL, et al., Methods, 2019). For Alas2_enhancer the enzymes used were DpnII>EcoRI while for St3gal6_TSS DpnII>Csp6I. For library preparation purified circular fragments were amplified by PCR with View-Point specific primers. Resulting amplicons were purified with AMPure XP beads (0.8×) and amplified using standard indexed Illumina primers as previously described (Krijger PHL, et al., Methods, 2019). Second-round PCR products were purified with PCR purification columns (Qiagen) and quantified by 2100 Bioanalyzer (Agilent, DNA 7500 kit). 4C library was prepared by equimolar mix of the samples. The library was cleaned-up by AMPure XP beads (0.8x) to remove PCR dimers before sequencing through Illumina MiSeq with a 75bp Single-End reads setup.