Source: Streptomyces werraensis 37
1000 u/ml Store at -20°C
Supplied in: 10 mM Tris-HCl (pH 7.6); 100 mM NaCl; 0,1 mM EDTA; 50% glycerol.
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.
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