ChIP-qPCR for Protein-DNA Interactions
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Corresponding Organization :
Other organizations : City of Hope, Stanford University, Cedars-Sinai Medical Center, University of California, Davis
Variable analysis
- Tissue type (mouse tissues)
- Formaldehyde treatment (1% formaldehyde for 15 min)
- Glycine treatment (0.150 M glycine for 10 min)
- Antibodies used for immunoprecipitation (AR and SOX2)
- Chromatin shearing size (200-500 bp)
- Chromatin DNA fragments analyzed by real-time qPCR
- PBS washing
- Cell lysis buffer composition (50 mM Tris-HCl (pH 8.0), 140 mM NaCl, 1 mM EDTA, 10% Glycerol, 0.5% NP-40, and 0.25% Triton X-100)
- Nuclear lysis buffer composition (10 mM Tris-HCl, pH 8.0, 1 mM EDTA, 0.5 mM EGTA, and 0.2% SDS)
- ChIP dilution buffer composition (0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl, pH 8.1, and 167 mM NaCl)
- Positive control: Magnetic protein G beads conjugated with AR or SOX2 antibody
- Negative control: Not explicitly mentioned
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