Church's Buffer for Northern and Southern Blot Hybridization
Church's buffer is commonly used for Northern and Southern blot hybridization.
Church's buffer composition
| Component | Target concentration |
|---|---|
| BSA | 1% (w/v) |
| EDTA | 1 mM |
| Sodium phosphate (pH 7.2) | 0.5 M |
| SDS | 7% (w/v) |
How to prepare Church's buffer
- Add approximately 800 mL of ultrapure or deionized water to a suitable vessel.
- Prepare a 0.5 M sodium phosphate system at pH 7.2 using both monobasic and dibasic sodium phosphate.
- Add BSA to 1% (w/v), EDTA to 1 mM, and SDS to 7% (w/v).
- Stir until all components are completely dissolved and the solution is uniform.
- Adjust the final volume with water as needed.
- Filter sterilize if required for the intended application.
Note: Use both monobasic and dibasic sodium phosphate to prepare the 0.5 M sodium phosphate component at pH 7.2.
Note: Adding SDS from an existing stock solution is often easier and faster than weighing SDS powder directly.
Reference: PNAS 81:1991-1995, April 1984.