GSM1480237: Smc1_ChIAPET_rep2; Mus musculus; OTHER
100 million murine ES cell were cross-linked by 1% formaldehyde at RT 10 mintues and were sonicated to solubilize the chromatin. The chromatin complexes contain DNA fragments with an average size of 300-800bp were immunoprecipitated by using murine SMC1a antibody. After end repair, the blunt end DNA were ligated with biotin labled half-linkers which containing flanking ECOP15i sites.  After eluted chromatin complexes from protein bead A, Proximity ligated was performed at extremely dilute conditions at 16 O/N. The purified DNA which digested by Ecop15I were prepared for sequencing according to a modified version of the pair end  DNA protocol. Fragmented DNA was end repaired and subjected to add "A" at 3' end, ligated with PE adaptor provided by Illumina. Amplified fragment 214bp (representing pair end genomic DNA 52bp, 42nt linker and  and ~120bp of adaptor sequence) were isolated by agarose gel electrophoresis and purified. sequencing was performed on Illumina Hi-Seq 2000 with PE 50 cycles. Linker A: 3'-GAC GAC AGG CTA TAG CGC CGG-5', 5'-CTG CTG TCC  GAT ATC GC-3'.  Linker B: 3'-GAC GAC AGT ATA TAG CGC CGG-5' 5'-CTG CTG TCA  TAT ATC GC-3'
GEO Accession
GSM1480237