162
edits
Changes
→Crosslinking, Lysis and Shearing of DNA
9. Resuspend each fresh or frozen pellet (containing 2 x 107 cells) on ice in 1 ml Farnham Lysis Buffer and mix gently by flicking the test tube. Briefly homogenize cells by running the cells through a 18-gauge needle ~10X.
''Note: This treatment breaks the cells while keeping the nuclei mostly intact.''
10. Collect the crude nuclear prep by centrifuging at 2,000 rpm at 4oC for 5 minutes.
