Caryophanon latum L
5’-A T C G A T-3’
3’-T A G C T A-5’
Reaction Temperature: 37°C
Prototype: ClaI
Inactivation Temperature (20 min): 65°C
Reaction Buffer:
1x Acet Buffer: 50 mM potassium acetate, 20 mM Tris-acetate (pH 7.5 at 37°C), 10 mM magnesium acetate, 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 potassium acetate, 20 mM Tris acetate (pH 7.5 at 37°C), 10 mM magnesium acetate, 1 mM dithiothreitol, 100 µg/ml bovine serum albumin and 1 µg of lambda DNA. Incubation is at 37°C for 1 hr in a reaction volume of 50 µl.
Storage Buffer: 50 mM Tris-HCl (pH 7.4 at 22°C), 200 mM KCl, 0.1 mM EDTA, 5 mM β-mercaptoethanol, 0.1% (v/v) Triton™X-100, 500 µg/ml bovine serum albumin and 50% (v/v) glycerol.
Storage Conditions: Store at –20°C.
Quality Control:
All preparations are assayed for contaminating endonuclease, 3'-exonuclease, 5'-exonuclease/5'-phosphatase, as well as non-specific single- and double-stranded DNase activities.