Ags I

Supplied in:

10 mM Tris-HCl (pH 7.5); 100 mM KCl; 0,1 mM EDTA;

200 µg/ml BSA; 7 mM  2-mercaptoethanol; 50% glycerol.

Reaction Conditions:

1X SEBuffer Y+BSA

Incubate at 37°C.

Warranty period for the enzyme storage at-20˚C is two years from the date of the last assay indicated on the enzyme vial.

1X SEBuffer Y  (pH 7.9 @ 25ºC)

33 mM Tris-Ac, 66 mM KAc, 10 mM MgAc, 1 mM DTT

Unit Definition:

One unit is defined as the amount of enzyme required to digest 1 µg of l DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Concentrated  enzymes are diluted to approximately

1000 units/ml with the buffer A [10 mM Tris-HCl (pH 7.6); 50 mM KCl; 0.1 mM EDTA; 1 mM DTT; 200 µg/ml BSA; 50% glycerol]  before determining their  activity.

To obtain 100% activity, BSA should be added to the 1X reaction mix to a final concentration of 100 µg/ml.

Quality Control Assays

Enzyme Properties

Activity in SEBuffers:

SEBuffer B       75-100%

SEBuffer G       50- 75%

SEBuffer O       10- 25%

SEBuffer W      10- 25%

SEBuffer Y           100%

SEBuffer ROSE    50%

When using a buffer other than the optimal (suppied) SEBuffer, it may be necessary to add more enzyme to achieve complete digestion. 

Heat Inactivation:

Yes   (65°C for 20 minutes) 

Reagents supplied with enzyme:

10X SEBuffer Y, BSA (10 mg/ml)