Abs I - CC↑TCGAGG / GGAGCT↓CC

Supplied in:

10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0,1 mM EDTA; 200 µg/ml BSA; 7 mM  2-mercaptoethanol; 0.05% Triton X-100; 50% glycerol.

Reaction Conditions:

1X SEBuffer Abs I.

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 Abs I  (pH 9.0 @ 25ºC)

10 mM Tris-HCl, 50 mM KCl, 10 mM MgCl2, 1 mM DTT

Unit Definition:

One unit is defined as the amount of enzyme required to digest 1 µg of pUC19SE/DriI in 1 hour at 37°C in a total reaction volume of 50 µl.

Quality Control Assays 

Ligation:

After 2-fold overdigestion with Abs I, ~90% of  DNA fragments can be ligated with T4 DNA Ligase and recut.

16-Hour Incubation:

A 50 µl reaction containing 1 µg of pUC19SE/DriI and 2 units of enzyme incubated for 16 hours resulted in the same pattern of DNA bands as a reaction incubated for 1 hour. A long incubation time may result in star activity.

Oligonucleotide Assay:

No detectable degradation of a single- and double-stranded oligonucleotide was observed after incubation with 1 units of enzyme for 3 hours.

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:

10XSEBuffer Y, BSA (10 mg/ml)