Southern blot denaturation solution recipe
Separates double-stranded DNA in the gel into single strands so the probe can bind: 1.5 M NaCl, 0.5 M NaOH.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| NaCl | 87.66 g | 1.5 M |
| NaOH | 20 g | 0.5 M |
| Water (distilled) | to final volume |
Steps
- Dissolve in about 80% of the final volume of water.
- Bring to the final volume with water.
- Store at room temperature.
- Soak the gel 2 × 15 min with shaking.
⚠ Safety Strongly alkaline — wear gloves and goggles.
FAQ
Can I skip neutralisation with alkaline transfer?
Yes — for alkaline transfer onto positively charged nylon in 0.4 M NaOH, neutralisation is skipped.
This is a widely used standard composition. Amounts depend on the hydrate form and purity of your reagents — check the formula weight (FW) on the bottle and follow your institution's safety rules and the manufacturers' instructions.