Reagents:
ChIP sonication Buffer (1% Triton X-100,0.1% Deoxycholate,50 mM Tris 8.1,150 mM NaCl,5 mM EDTA):
10 ml 10% Triton X-100
1 ml 10% Deoxycholate
5 ml 1 M Tris-Cl pH 8.1
1 ml 0.5 M EDTA
3 ml 5 M NaCl
80 ml Water
Just before use,add 10μl Aprotinin,10μl Leupeptin,and 5μl PMSF to each 10 ml.
High Salt Wash Buffer (1% Triton X-100,0.1% Deoxycholate,50 mM Tris-8.1,500 mM NaCl,5 mM EDTA)
10 ml 10% Triton X-100
1 ml 10% Deoxycholate
5 ml 1M Tris-8.1
1 ml 0.5M EDTA
10 ml 5M NaCl
73 ml Water
(LiCl Immune Complex Wash Buffer)
25 ml 1mliCl
5 ml 10% IGEPAL
5 ml 10% Deoxycholate
1 ml 1M Tris-8.1
200μl 0.5M EDTA
64 ml Water
Protease inhibitors (add 10μl of each to 10 ml of PBS or sonication buffer)
Leupeptin 2 mg/ml in water
Aprotinin 2 mg/ml in water
PMSF 0.2 M
5 M NaCl
1X TE Buffer (10mM Tris,8.1,1 mM EDTA)
0.5 M EDTA
1 M Tris-Cl,pH 6.8
Protein A/G Agarose (Santa Cruz #sc-2003)
Proteinase K (19 mg/ml,Boehringer Mannheim # 1964372)
10X proteinase K buffer
Elution Buffer (1% SDS,0.1 M NaHCO3,0.01 mg/ml Herring sperm DNA)
10 mg/ml Herring Sperm DNA
37% Formaldehyde (ACS reagent grade)
1.25 M glycine