Hosted @RenLab

Protocols used in ChIP-on-chip experiments

Solutions used in ChIP-on-Chip experiments:

1. 2% of BSA: 1 g of BSA in 50 ml of Mili-Q water
2. Cross-linking solution: 11% formaldehyde, 0.1M NaCl, 1mM Na-EDTA, 0.5mM Na-EGTA, 50 mM Hepes, pH 8.0
3. Lysis Buffer 1: 0.05 M Hepes-KOH, pH 7.5, 0.14 M NaCl, 1mM EDTA, 10% glycerol, 0.5% NP-40, 0.25% Triton X-100, with protease inhibitor cocktail (Roche Applied Science)
4. Lysis Buffer 2: 0.2 M NaCl, 1mM EDTA, 0.5 mM EGTA, 10 mM Tris pH 8, protease inhibitor cocktail
5. Lysis Buffer 3: 1mM EDTA, 0.5 mM EGTA, 10 mM Tris-HCl pH 8, protease inhibitor cocktail
6. Proteinase K stock solution: 20 mg/ml proteinase K (Sigma) 50 mM Tris-HCl pH 8.0, 1.5 mM calcium Acetate
7. RIPA buffer: 50 mM Hepes, pH 8.0, 1mM EDTA, 0.7% DOC, 1% NP-40, 0.5M LiCl, protease inhibitor cocktail
8. Elution buffer: 10 mM Tris pH 8, 1 mM EDTA, 1% SDS
9. Array Pre-hybridization Solution: 2XSSC/0.05%SDS/0.2%BSA
10. Hybridization buffer 1: 2.2x SSC, 0.22% SDS
11. Hybridization buffer 2: 70% formamide, 3XSSC, 14.3% dextran sulfate