This is a wash buffer for washing samples for in situ hybridization and similar procedures.
Optional add % (vol/vol) of formamide, e.g. 10% formamide if needed. If you aim is to wash away formamide in tissue don't use formamide in the wash buffer.
DEPC-water is recommended if cDNA is not synthesized yet. Post cDNA nuclease-free water suffices.
| Item | Item article number | Distributor | Vendor |
|---|---|---|---|
| DEPC-water | 10419224 |
ThermoFisher | ThermoFisher |
| 20x SSC (get from stock room) | 10085664 | ThermoFisher | ThermoFisher |
| Tween-20 | Merck | SigmaAldrich | |
| Hi-D formamide | 4311320 | ThermoFisher | ThermoFisher |
First make a stock of 10% (vol/vol) Tween-20 in a 15 mL Falcon tube. Store for 6-12 months at room temp. Cut the tip of the 1 mL pipette to more easily pipette the viscous Tween-20.
| Reagent | Amount | Final concentration |
|---|---|---|
| Nuclease-free water | 4.5 mL | |
| Tween-20 | 500 µL | 10% (vol/vol) |
| TOTAL: | 5 mL |
Vortex heavily and wait until the bubbles/foam have disappeared.
Next make the 50 mL wash buffer in a 50 mL Falcon tube. Throw away after 6 months. Store in room temperature.
| Reagent | Amount | Final concentration |
|---|---|---|
| 20x SSC | 5 mL | 2x |
| DEPC-water | 39.5 mL | |
| Formamide (optional, leave out and use water instead) | 5 mL | 10% (vol/vol) |
| 10% (vol/vol) Tween-20 | 500 µL | 0.1% (vol/vol) |
| TOTAL: | 50 mL |