Chromatin Immunoprecipitation: Difference between revisions
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===== 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 | * Set water bath to 65°C for use later. | ||
29. Make Elution Buffer for all IP tubes as well as all Input tubes | 29. Make Elution Buffer for all IP tubes as well as all Input tubes. | ||
* For each tube, prepare 200 μL of elution buffer as follows: 10 μL 20% SDS, 20 μL 1 M NaHCO3 and 170 μL sterile, distilled water. | * For each tube, prepare 200 μL of elution buffer as follows: 10 μL 20% SDS, 20 μL 1 M NaHCO3 and 170 μL sterile, distilled water. | ||
* OR can make this way [[ChIP Elution Buffer]] | |||
* Alternatively, make a large volume to accommodate all tubes. For example, if there are 10 tubes mix together 105 μL 20% SDS, 210 μL 1M NaHCO3 and 1.785 mL sterile, distilled water. | * Alternatively, make a large volume to accommodate all tubes. For example, if there are 10 tubes mix together 105 μL 20% SDS, 210 μL 1M NaHCO3 and 1.785 mL sterile, distilled water. | ||
30. For Input tubes | 30. For Input tubes, add 200 μL of Elution Buffer and set aside at room temperature. | ||
31. Add 100 μL of Elution Buffer to each tube containing the antibody/agarose complex. Mix by flicking tube gently. | 31. Add 100 μL of Elution Buffer to each tube containing the antibody/agarose complex. Mix by flicking tube gently. | ||