Chromatin Immunoprecipitation: Difference between revisions
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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. | 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. | ''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. | 10. Collect the crude nuclear prep by centrifuging at 2,000 rpm at 4oC for 5 minutes. | ||