Hybe Solution
- 50% Formamide (Amresco, ultrapure)
- 5x SSC
- 100 ug/mL salmon sperm DNA
- 50 ug/mL heparin
- 0.1% Tween-20
| Reagent | For 250 mL stock |
| Formamide | 125 mL |
| 20x SSC | 62.5 mL |
| Heparin 10 mg/mL lab aliquot | 1.25 mL |
| 10% Tween-20 | 2.50 mL |
| ssDNA 10 mg/mL lab aliquot | 2.50 mL |
| ddH2O | 56.25 mL |
SSC = Na3CitrateX2H2O
PBT
| Reagent | For 1 L stock |
| PBS 10x | 100 mL |
| 10% Tween-20 | 10 mL |
| ddH2O | 890 mL |
Staining Buffer
| Reagent | For 50 mL | For 1 mL |
| NaCl 5M | 1 mL | 20 uL |
| MgCl 1M | 2.5 mL | 50 uL |
| Tris pH 9.5 1M | 5 mL | 100 uL |
| 10% Tween-20 | 0.5 mL | 10 uL |
| ddH2O | 41 mL | 820 uL |
Staining Solution
To 1 mL of staining buffer, add 9 or 13.5 uL NBT and 17.5 or 7 uL BCIP
- If lab stock (75 mg/mL) add 9 uL NBT
- If store bought (50 mg/mL) add 13.5 uL NBT
- If lab stock (20 mg/mL) add 17.5 uL BCIP
- If store bought (50 mg/mL) add 7 uL BCIP