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Specifications

5'-G G N^N C C-3'
3'-C C N^N G G-5'
Catalog #	B2960
Concentration	10u/ul
Source	Bacillus species RJ3–212
Buffer	33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C
Diluent Buffer	10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol, or Storage Buffer.
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 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BspLI.
Ligation/Recutting Assay	After 50-fold overdigestion (3u/ug DNA x 17 hours) with BspLI, approximately 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.78uM. 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.
Methylation Effects	BspLI cleaves the hemi-methylated sequence GGNNCm5C. Cleavage is blocked when both strands are methylated at positions: 5… GGNNCm5C…3' 3'…m5CCNNG G…5'. Overlapping Dcm or CG methylation may influence DNA cleavage.
Stability during Prolonged Incubation	A minimum of 0.3units of enzyme is required for complete digestion of 1ug of DNA in 16 hours at 37°C.
Thermal Inactivation	Enzyme is inactivated by incubation at 65°C for 20min.
Number of Recognition Sites in DNA
Lambda	82
PhiX174	6
M13mp18/19	18
pBR322	24
pUC18/19	11
pUC57	12
pTZ19R/U	13
pBluescriptIIKS(-/+)	15
pBluescriptIISK(-/+)	15
pACYC177	10
pACYC184	23
Unit Definition	One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.