EB buffer (10 mM Tris-HCl pH 8.5) recipe
An EDTA-free buffer for eluting DNA from columns and storing it without interfering with sequencing or enzymes.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| 1 M Tris-HCl (pH 8.5) | 1 mL | 10 mM |
| Water (distilled) | to final volume |
Steps
- Add the Tris-HCl stock to water and mix.
- Autoclave (121 °C, 20 min) and store at room temperature.
Good to know
- Its stable pH protects DNA from acid hydrolysis, unlike water.
FAQ
Why not TE?
TE is better for long-term storage, but EDTA chelates Mg²⁺ and can inhibit PCR, sequencing and enzymes.
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.