Chromatin Immunoprecipitation: Difference between revisions
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14. Snap-freeze the sample in liquid nitrogen and store at -80°C, or do not freeze and continue with the immunoprecipitation steps below. | 14. Snap-freeze the sample in liquid nitrogen and store at -80°C, or do not freeze and continue with the immunoprecipitation steps below. | ||
===Immunoprecipitation=== | ===Immunoprecipitation===(taken from Millipore EZ ChIP protocol) | ||
''Perform all steps in an ice bucket or in the cold room at 4°C.'' | ''Perform all steps in an ice bucket or in the cold room at 4°C.'' | ||
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* LiCl Immune Complex Wash Buffer (Catalog # 20-156), one wash | * LiCl Immune Complex Wash Buffer (Catalog # 20-156), one wash | ||
* TE Buffer (Catalog # 20-157), two washes | * TE Buffer (Catalog # 20-157), two washes | ||
==== Elution of Protein/DNA Complexes ==== | |||
Prior to starting this section: | ===== Prior to starting this section: ===== | ||
• Bring 1 M NaHCO3 to room temperature. A precipitate may be observed but will go into solution once room temperature is achieved. The 1 M NaHCO3 can be vortexed. | • Bring 1 M NaHCO3 to room temperature. A precipitate may be observed but will go into solution once room temperature is achieved. The 1 M NaHCO3 can be vortexed. | ||
• Set water bath to 65°C for use in Section E. | • Set water bath to 65°C for use in Section E. | ||