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Operator’s
Manual
Rev
A
4/30/12
pg.
73
Appendix
A
–
Reagents
(AOAC
991.43,
AACC
32
‐
07.01,
NMKL
129,
2003)
Use deionized water throughout.
(a)
Ethanol 95%
.
(b)
Ethanol 78%
—Place 821 ml 95% ethanol into 1 L volumetric flask, dilute to volume with H
2
O.
(c)
Heat-stable
α
-amylase solution
—Dilute 1.25 ml of Megazyme amylase to 25 ml with a solution of 25% Glycerol and
0.02% Sodium Azide in Distilled Water.
(d)
Protease
—Dilute 2.5 ml of Megazyme protease to 25 ml with a solution of 25% Glycerol and 0.02% Sodium Azide in
Distilled Water.
(e)
Amyloglucosidase (AMG) solution
—Dilute 5.0 ml of Megazyme AMG to 25 ml with a solution of 25% Glycerol and
0.02% Sodium Azide in Distilled Water.
(f)
Diatomaceous earth (DE)
—(ANKOM XTC, or equivalent).
(g)
MES
—2-(
N
-Morpholino)ethanesulfonic acid (No. M-8250, Sigma Chemical Co., or equivalent).
(h)
TRIS
—Tris(hydroxymethyl)aminomethane (No. T-1503, Sigma Chemical Co., or equivalent).
(i)
MES-TRIS buffer solution
—0.05M MES, 0.05M TRIS, pH 8.2 at 24°C. Dissolve 19.52 g MES and 12.2 g TRIS in 1.7 L
H
2
O. Adjust pH to 8.2 with 6
N
NaOH, and dilute to 2 L with H
2
O. (
Note:
It is important to adjust pH to 8.2 at 24°C.
However, if buffer temperature is 20°C, adjust pH to 8.3; if temperature is 28°C, adjust pH to 8.1. For deviations between
20 and 28°C, adjust by interpolation.)
(j)
Hydrochloric acid solution
—0.561
N
. Add 93.5 ml 6
N
HCl to ca 700 mL H
2
O in 1 L volumetric flask. Dilute to 1 L with
H
2
O.
(k)
Acetone
—reagent grade.
(l)
Sodium Azide
—reagent grade.
Summary of Contents for TDF
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