BalI
(Brevibacterium albidum)
5’-T G G C C A-3’
3’-A C C G G T-5’
Reaction Temperature: 37°C
Prototype: BalI
Inactivation Temperature (20 min): 65°C
Reaction Buffer:
1x Acet Buffer: 20 mM Tris-acetate (pH 7.5 at 37°C), 10 mM magnesium acetate, 50 mM potassium 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. Total reaction volume is 30 µl.
Assay Conditions: 20 mM Tris-acetate (pH 7.5 at 37°C), 10 mM magnesium acetate, 50 mM potassium 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 30 µl.
Storage Buffer: 10 mM Tris-HCl (pH 7.5 at 22°C), 1 mM dithiothreitol, 50 mM KCl, 0,1 mM EDTA, 200 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, and nonspecific single- and double-stranded DNase activities.