Supplied in:

10 mM Tris-HCl (pH 7.6); 100 mM NaCl; 0,1 mM EDTA; 50% glycerol.

Reaction Conditions:

1XSEBuffer Y

Incubate at 55 °C.

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

1XSEBuffer Y:

33 mM Tris-Ac (pH 7.9 @ 25ºC), 66 mM KAc, 10 mM MgAc, 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`-CGAGTTTATAGCTGGGCCCAAC-3` 

3`-GCTCAAATATCGACCCGGGTTG-5` in 1 hour at 55ºC in a total reaction volume or 20 µl

Quality Control Assays

Ligation:

After 5-fold overdigestion with Set I, approximately 50% of DNA fragments can be ligated with T4 DNA Ligase and  recut.

*SetI cleaves a canonical site and several other sites with a weaker activity. In the case of long incubation with SetI DNA can be digested to small oligos.

Oligonucleotide Assay:

No detectable degradation of a single- and double-stranded oligonucleotide was observed after incubation with 5 units of enzyme for 3 hours.

Enzyme Properties

Activity in SEBuffers:

SEBuffer B         25- 50%

SEBuffer G         25- 50%

SEBuffer O         75-100%

SEBuffer W        75-100%

SEBuffer Y               100%

SEBuffer ROSE        100%

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   (80°C for 20 minutes)

Reagents supplied with enzyme:

10X SEBuffer Y