ChIP/Liver harvest

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Materials)
Line 4: Line 4:
==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 14:08, 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)

Contact

Personal tools