buffers.md

Buffers and Reagents

• Tris (1 M, pH 7.5)
• TAE 50x
• EDTA 0.5 M, pH 8.0
• Ethidium Bromide 10 mg/ml

Tris (1 M, pH 7.5)

Tris recipe for buffering V. fischeri growth media.

Amount	Reagent
127 g	Tris-HCl
23.6 g	Tris base

Stir until dissolved, adjust to pH 7.5, then adjust volume to 1L with dI H₂O.

Filter-sterilize and store at room temperature.

TAE 50x

242 g Tris Base 57.1 mL Acetic acid, glacial 100 ml EDTA (0.5 M)

Adjust volume to 1L with dI H2O.

Store at room temperature.

EDTA 0.5 M, pH 8.0

ethylenediaminetetraacetic acid

Dissolve 186.1 g of disodium EDTA in 800 mL water.
Adjust pH with NaOH. Adjust volume to 1L with dI H2O. Aliquot and autoclave.

Note: EDTA will not go into solution until the pH is approximately 8.0.

Ethidium Bromide 10 mg/ml

Add 250 mg of ethidium bromide to 25 mL of dH2O in a conical tube (amber, if available).

If not amber, wrap in aluminum foil. Invert on a wrist shaker for several hours to ensure that the dye has dissolved.

Store at room temperature.