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 B.

Incubate at 50°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 B  (pH 7.6 @ 25ºC)

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

Unit Definition:

One unit is defined as the amount of enzyme required to cleave 1 pmol of the double stranded oligonucleotide with the following structure

5'-CGAGTTCA^TAGCTGGGCCCAAC-3'

3'-GCTCAAGT^ATCGACCCGGGTTG-5'

in 1 hour at 50°C in a total reaction volume of 20 μl.

Quality Control Assays

Ligation: After 3-fold overdigestion with Fai I, ~50% of pUC19 DNA fragments can be ligated with T4 DNA Ligase and recut.

Oligonucleotide Assay:

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

Enzyme Properties 

Activity in SEBuffers:

SEBuffer B         100%

SEBuffer G     50-75%

SEBuffer O     10-25%

SEBuffer W     25-50%

SEBuffer Y      25-50%

SEBuffer ROSE  100%

Heat Inactivation:

Yes   (80°C for 20 minutes)

Reagents supplied with enzyme:

10X SEBuffer B