Hoefer HE99X - Manual (Page 19)

2. 10X Tris-phosphate-EDTA (TPE) stock buffer a
(0.89 M Tris, 0.89 M phosphoric acid, 20 mM EDTA,
pH ≈8.1, 1000 ml)
Tris base (FW 121.1)
0.89 M 108.0 g
Phosphoric acid (85%) 0.23 M 15.5 ml
EDTA solution 0.02 M 40.0 ml
(0.5 M, pH 8.0, solution 4)
Deionized H
2
O to 1000.0 ml
Stir. Do not adjust pH. Dilute to 1X, to yield 89 mM
Tris base, 23 mM phosphoric acid, and 2 mM EDTA.
3. 10X Tris-acetate-EDTA (TAE) stock buffer a
(0.4 M Tris, 0.2 M acetic acid, 10 mM EDTA,
pH ≈8.4, 1000 ml)
Tris base (FW 121.1)
0.40 M 48.4 g
Acetic acid (99.5%) 0.20 M 11.4 ml
EDTA solution
(0.5 M, pH 8.0, solution 4)
0.01 M 20.0 ml
Deionized H
2
O to 1000.0 ml
Stir. Do not adjust pH. Dilute to 1X before use to
yield 40 mM Tris base, 20 mM acetic acid,
and 1 mM EDTA.
4. EDTA solution
(ethylenediamine tetraacetic acid)
b
(0.5 M, pH 8.0, 100 ml)
Na
2
EDTA·2H
2
O, (FW 372.2) 0.5 M 18.6 g
Deionized H
2
O to 70.0 ml
NaOH (10 M) to pH 8.0 ≈5.0 ml
Deionized H
2
O to 100.0 ml
a Sambrook J. and Russell, D.W., (2001) Molecular Cloning: A Labora-
tory Manual, A1.17
b Current Protocols in Molecular Biology (1993) A.2.1
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