Unit Definition

One unit is defined as the amount of enzyme required to protect 1 μg λ DNA in 1 hour at 65°C in a total reaction volume of 20 μl against cleavage by TaqI restriction endonuclease.

Reaction Conditions

1X rCutSmart™ Buffer
Supplement with 80 µM S-adenosylmethionine (SAM)
Incubate at 65°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Control Protection Assay

Protection Assay Conditions:
TaqI Methyltransferase is incubated with 1 µg λ DNA in 20 µl 1X rCutSmart Buffer and 80 µM S-adenosylmethionine, for 1 hour at 65°C. The extent of protection by TaqI Methyltransferase is determined by the addition of 30 µl 1X rCutSmart Buffer and 10 units of TaqI restriction endonuclease. Incubation for 15 minutes at 65°C is followed by analysis on an agarose gel.

Storage Buffer

10 mM Tris-HCl
100 mM NaCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Molecular Weight

Theoretical: 47845 daltons