ChIP/Liver harvest
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==Materials== | ==Materials== | ||
| - | + | '''Buffer A (100mL total)'''<br> | |
| + | * 91mL of H2O<br> | ||
| + | * 1.5mL of 1M Hepes<br> | ||
| + | * 2mL of 3M KCl<br> | ||
| + | * 300µL of 5M NaCl<br> | ||
| + | * 40µL of 0.5M EDTA<br> | ||
| + | * 100µL of 0.5M EGTA<br> | ||
| + | * 16.2µL of 2.1M Sucrose<br> | ||
| + | * 7.5µL of 2M BME<br> | ||
| + | *5mL of 2.5M Glycine (to make 2.5M stock- 187.5 g/L)<br> | ||
| + | **A stock of Liver Buffer A can be made without BME or Glycine and stored at room temp. Add BME and Glycine before use. | ||
| + | <br> | ||
| + | '''Buffer B (100mL total)'''<br> | ||
| + | * 1.5mL of 1M Hepes<br> | ||
| + | * 2mL of 3M KCl<br> | ||
| + | * 300µm of 5M NaCl<br> | ||
| + | * 20µm of 0.5M EDTA<br> | ||
| + | * 20µm of 0.5M EGTA<br> | ||
| + | * 100mL of 2.1M Sucrose<br> | ||
| + | <br> | ||
| + | '''Other''' | ||
| + | * PBS<br> | ||
| + | * PBS/1% Formaldehyde<br> | ||
| + | * Avertin (Anesthesia) | ||
==Procedure== | ==Procedure== | ||
Revision as of 15:08, 6 December 2006
Contents |
Overview
This is a protocol for harvesting mouse liver tissue for chromatin immunoprecipitation.
Materials
Buffer A (100mL total)
- 91mL of H2O
- 1.5mL of 1M Hepes
- 2mL of 3M KCl
- 300µL of 5M NaCl
- 40µL of 0.5M EDTA
- 100µL of 0.5M EGTA
- 16.2µL of 2.1M Sucrose
- 7.5µL of 2M BME
- 5mL of 2.5M Glycine (to make 2.5M stock- 187.5 g/L)
- A stock of Liver Buffer A can be made without BME or Glycine and stored at room temp. Add BME and Glycine before use.
Buffer B (100mL total)
- 1.5mL of 1M Hepes
- 2mL of 3M KCl
- 300µm of 5M NaCl
- 20µm of 0.5M EDTA
- 20µm of 0.5M EGTA
- 100mL of 2.1M Sucrose
Other
- PBS
- PBS/1% Formaldehyde
- Avertin (Anesthesia)
Procedure
Notes
- Please contribute user notes!
References
- (Young Lab)
