GSM1689153: Smc1_ChIAPET_rep1; Homo sapiens; OTHER
Whole cell extracts were sonicated to solubilize the chromatin. The chromatin extracts containing DNA fragments with an average size of 300-700 bp were immunoprecipitated using SMC1 antibody. Immunoprecipitated DNA was end-repaired, A-tailed, and proximity ligation was performed in the presence of a bridge-linker sequence. Unligated DNA was digested with exonuclease treatment followed by RNAse A, and Proteinase K treatment. DNA was then phenol:chloroform:isoamyl alcohol preciptated and subjected to library preperation. Purified immunoprecipitated DNA were prepared for sequencing according to the Illumina Nextera Tagmentation protocol. Tagmented DNA was subjected to 12 cycles of LM-PCR using oligos provided by Illumina. Amplified fragments between 300 and 500bp (representing shear fragments between 200 and 400 nt in length and ~100bp of primer sequence) were isolated by agarose gel electrophoresis and purified.
GEO Accession
GSM1689153