HotSHOT neutralisation reagent recipe

Base: 1 L · pH 5.0 · Updated 2026-10-01

Added in an equal volume after HotSHOT alkaline lysis to bring the pH back: 40 mM Tris-HCl, pH 5.0.

🧮 Volume calculator
ComponentAmount Final concentration (1X)
Tris-HCl (Tris hydrochloride)630 mg40 mM
Water (distilled)to final volume

Steps

  1. Dissolve in about 80% of the final volume of water.
  2. Check pH 5.0 and adjust with HCl if needed.
  3. Bring to the final volume with water.
  4. Store at room temperature.

FAQ

Can I make it from Tris base?

Yes — dissolve 4.85 g/L Tris base and titrate to pH 5.0 with HCl.

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.