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→Reverse Crosslinks of Protein/DNA Complexes to Free DNA
=== Reverse Crosslinks of Protein/DNA Complexes to Free DNA===
36. To all tubes (IPs and Inputs) add 8 μL 5 M NaCl and incubate at 65°C for 4-5 hours or '''overnight ''' to reverse the DNA – Protein crosslinks. After this step the sample can be stored at -20°C and the protocol continued the next day.
37. To all tubes, add 1 μL of RNase A and incubate for '''30 minutes ''' at 37°C.
38. Add 4 μL 0.5M EDTA, 8 μL 1M Tris-HCl and 1 μL Proteinase K to each tube and incubate at 45°C for
'''1-2 hours'''.
==== Purification of ChIP DNA ====