Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to protect 1 μg λ DNA in 1 hour at 37°C in a total reaction volume of 10 μl against cleavage by EcoRI restriction endonuclease Reaction Conditions
1X rCutSmart™ Buffer
Supplement with 640 µM S-adenosylmethionine (SAM)
Incubate at 37°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
One unit of EcoRI Methyltransferase is incubated with 1 µg of λ DNA in 10 µl 1X rCutSmart supplemented with 640 µM SAM for one hour at 37°C followed by 15 min inactivation at 65°C. The extent of protection by EcoRI Methyltransferase is determined by the addition of 10 units of EcoRI restriction endonuclease in 40 µl of 1X rCutSmart Buffer followed by incubation at 37°C for 30 minutes. Results are analyzed on agarose gel. Storage Buffer
100 mM KPO4
200 mM NaCl
10 mM β-ME
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C
Heat Inactivation
65°C for 15 minutes Molecular Weight
Theoretical: 37911 daltons