Mirmira Lab ChIP Protocol

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 10 ul Aprotinin, 10 ul Leupeptin, and 5 ul 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 1M LiCl

 

 

5 ml 10% IGEPAL

 

 

5 ml 10% Deoxycholate

 

 

1 ml 1M Tris-8.1

 

 

200 ul 0.5M EDTA

 

 

64 ml Water

 

 

Protease inhibitors (add 10 ul of each to 10 ml of PBS or sonication buffer)