DdeI
(Desulfovibrio desulfuricans)
5’-C T N A G-3’
3’-G A N T C-5’
Reaction Temperature: 37°C
Prototype: DdeI
Inactivation Temperature (20 min): 65°C
Reaction Buffer:
1x High Buffer: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl,1 mM dithiothreitol, 100 µg/ml bovine serum albumin.
Unit Definition: One unit is the amount of enzyme required to completely digest 1 µg of lambda DNA in 1 hr in a total reaction volume of 50 µl.
Assay Conditions: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, 1 mM dithiothreitol, 100 µg/ml bovine serum albumin, 1 µg of lambda DNA. Incubation is at 37°C for 1 hr in a reaction volume of 50 µl.
Storage Conditions: Store at –20°C.
Quality Control: All preparations are assayed for contaminating endonuclease, 3'-exonuclease, 5'-exonuclease/5'-phosphatase, for nickase as well as for nonspecific single- and double-stranded DNase activities.