ChIP assay was performed as described previously 40 (link). In brief, cells were crosslinked with 1% formaldehyde and the crosslinked chromatin was fragmented to 200-500 bp by sonication. The appropriate amounts of lysates were incubated with Magna ChIP Protein G Magnetic beads (Millipore) and specific antibodies (5 μg) at 4 °C overnight with gentle rocking. The ChIP complexes were eluted and analyzed by qRT-PCR (primer sequences listed in Table S1). Fold enrichment was calculated by the ΔΔCt method.
Other organizations :
National Tsing Hua University, Taichung Veterans General Hospital, Chung Shan Medical University, Chang Gung University, National Health Research Institutes, Institute of Information Science, Academia Sinica, Chang Gung Memorial Hospital, Taipei Medical University, University of California, Davis
Fragmenting the crosslinked chromatin to 200-500 bp by sonication
Incubating the lysates with Magna ChIP Protein G Magnetic beads and specific antibodies (5 μg) at 4 °C overnight with gentle rocking
dependent variables
ChIP complexes eluted and analyzed by qRT-PCR
control variables
Crosslinking and chromatin fragmentation conditions
Antibody concentration and incubation conditions
controls
Not explicitly mentioned
Not explicitly mentioned
Annotations
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