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Specifications

5'-G C A G C (N)8 ^-3'
3'-C G T C G (N)12^-5'
Catalog #	B2920
Concentration	3u/ul
Source	Bacillus stearothermophilus Ra 3–212
Buffer	50mM Tris-HCl (pH 7.5), 2mM MgCl2,100mM NaCl and 0.1mg/ml BSA. Incubate at 65°C. (Incubation at 37°C results in 10% activity.)
Diluent Buffer	10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods–the Storage Buffer should be used.
Storage Buffer	10mM Tris-HCl (pH 7.4 at 25°C), 200mM NaCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion Assay	No detectable change in the specific fragmentation pattern is observed after 32-fold overdigestion (2u/ug pBR322 DNA x 16 hours) with BseXI (see Star Activity).
Ligation/Recutting Assay	After 10-fold overdigestion (0.6u/ug DNA x 17 hours) with BseXI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.98uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) Assay	No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.
Star Activity	A large excess of enzyme (4u/ug DNA x 16 hours) may result in star activity.
Stability during Prolonged Incubation	A minimum of 0.3units of enzyme is required for complete digestion of 1ug of pBR322 DNA in 16 hours at 37°C.
Thermal Inactivation	Enzyme is inactivated by incubation at 80°C for 20min.
Number of Recognition Sites in DNA
Lambda	199
PhiX174	14
M13mp18/19	10
pBR322	21
pUC18/19	12
pUC57	12
pTZ19R/U	12
pBluescriptIIKS(-/+)	13
pBluescriptIISK(-/+)	13
pACYC177	7
pACYC184	15
Unit Definition	One unit is defined as the amount of enzyme required to digest 1ug of pBR322 DNA in 1 hour at 65°C in 50ul of assay buffer.