New England Biolabs: Reagents for the Life Sciences Industry
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Glycosidases Double Digest Chart

NEB's glycosidases are provided with 10X buffer to ensure optimal activity. Using more than one glycosidase simultaneously is a common timesaving procedure. Selecting the best buffer to provide reaction conditions that optimize enzyme activity is an important consideration. The following chart will help in selecting the optimal conditions for double digests.

NEB # P0734 P0732 P0721 P0702
P0703
P0733 P0724 P0731 P0726 P0730 P0729 P0727 P0720 P0728 P0704
P0705
P0704
P0705
G7 G7 G7 S G7 G7 G7 G7 G7 G7 G7 G7  G7 G7
P0702
P0703
S S S G5 G5 S S S S S S G5 S S
P0733 G7 G7 G7 G5 G7 G7 G7 G7 G7 G7 G7 G7 G7 G7
P0726 G6 G6 G6 S G7 G6 G6 G2 G6 G6 G6 G6 G6 G7
P0730 G6 G6 G6 S G7 G6 G6 G6 G4 G6 G6 G6 G6 G7
P0731 G6 G6 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G6 G7
P0732 G6 G1 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G6 G7
P0734 G7 G6 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G6 G7
P0721 G6 G6 G2 S G7 G6 G6 G6 G2 G6 G6 G6 G6 G7
P0729 G6 G6 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G6 G7
P0727 G6 G6 G6 S G7 G6 G6 G6 G6 G6 G2 G6 G6 G7
P0720 G6 G6 G6 G5 G7 G6 G6 G6 G6 G6 G6 G1 G6 G7
P0728 G6 G6 G6 S G7 G6 G6 G6 G6 G6 G6 G6 G4 G7
P0724 G6 G6 G6 S G7 G4 G6 G6 G6 G6 G6 G6 G6 G7

Notes

P0702 = Endo H
P0703 = Endo Hf
P0704 = PNGaseF
P0705 = PNGaseF (Glycerol Free)
P0720 = α2-3,6,8 Neuraminidase
P0721 = β-N-Acetylhexosaminidasef
P0724 = α1-2 Fucosidase
P0726 = β1-3 Galactosidase
P0727 = α1-6 Mannosidase
P0728 = α2-3 Neuraminidase
P0729 = α1-2,3 Mannosidase
P0730 = β(1-4) Galactosidase
P0731 = α(1-3,6) Galactosidase
P0732 = β-N-Acetylglucosaminidase
P0733 = O-Glycosidase
P0734 = α-N-Acetylgalactosaminidase

S = Sequential digest under denaturing conditions
(Heat kill and sequential digest with appropriate buffer for each enzyme)
If BSA is required for one enzyme, include it in the double digest.

Reaction Buffer Compositions

1X G1 Reaction Buffer:
50 mM Sodium Citrate (pH 6.0 @ 25°C).

1X G2 Reaction Buffer:
50 mM Sodium Citrate (pH 4.5 @ 25°C).

1X G4 Reaction Buffer:
50 mM Sodium Citrate (pH 6.0 @ 25°C)
and 100 mM NaCl.
1X G5 Reaction Buffer:
50 mM Sodium Citrate (pH 5.5 @ 25°C).

1X G6 Reaction Buffer:
50 mM Sodium Acetate (pH 5.5 @ 25°C)
and 5 mM CaCl2.

1X G7 Reaction Buffer:
50 mM Sodium Phosphate (pH 7.5 @ 25°C).