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.
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
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
After 2-fold overdigestion with Abs I, ~90% of DNA fragments can be ligated with T4 DNA Ligase and recut.
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.
No detectable degradation of a single- and double-stranded oligonucleotide was observed after incubation with 1 units of enzyme for 3 hours.
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.
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