Chromatin Immunoprecipitation: Difference between revisions
→Crosslinking, Lysis and Shearing of DNA: specified tube for sonicaiton |
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10. Collect the crude nuclear prep by centrifuging at 2,000 rpm at 4°C for 5 minutes. | 10. Collect the crude nuclear prep by centrifuging at 2,000 rpm at 4°C for 5 minutes. | ||
11. Resuspend pellet to 1 ml with RIPA Buffer (Do not vortex the tubes and try to avoid bubbles. Bubbles will cause popping and loss of samples during sonication). | 11. Resuspend pellet to 1 ml with RIPA Buffer in a 15 mL falcon tube (Do not vortex the tubes and try to avoid bubbles. Bubbles will cause popping and loss of samples during sonication). | ||
12. Using the Sonics VibraCell Sonicator, sonicate each 1.0 ml ChIP sample on ice, in a cold room, at Power Output 5 watts 6 times for 30 seconds each | 12. Using the Sonics VibraCell Sonicator, sonicate each 1.0 ml ChIP sample on ice, in a cold room, at Power Output 5 watts 6 times for 30 seconds each | ||