GSM5462293: Atria_rep1_CAV1_GATC_CATG; Mus musculus; OTHER
Heart were dissected and atria tissue were micro-dissected. Pools of 20 atria were dissociated to obtain single cells preparations with collagenase. Chromatin was fixed with 2% formaldehyde for 10 min at room temperature and the reaction was quenched on ice with glycine. After fixation, pools were lysed in 50mM Tris-HCl pH7.5, 150 mM NaCl, 5 mM EDTA, 0.5% NP-40, 1% TX-100 and 1x complete protease inhibitor (Roche), digested first with DpnII (New England Biolabs, Cat. No. R0543M), ligated with T4 DNA ligase (Promega) followed by either NlaIII (New England Biolabs, Cat. No. R0125L) or Csp6I (Fermentas, cat.no. ER0211) digestion, depending on the initial design of the 4C viewpoint, and re-ligated with T4 DNA ligase (Promega). 500ng of 4C templates were amplified using Expand Long Template Polymerase enzyme (Roche) and inverse PCR primers flanked with barcodes allowing multiplexing. The Illumina adaptors used were 5'-AATGATACGGCGACCACCGAACACTCTTTCCCTACACGACGCTCTTCCGATCT-3' for the reading primers and 5'-CAAGCAGAAGACGGCATACGA-3' for the non-reading. Specific primer information for each viewpoint is displayed in the related publication.