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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BspPI (BinI)

BspPI (BinI)

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Specifications

5'-G G A T C (N)4^-3'
3'-C C T A G (N)5^-5'
Catalog #B2965
Concentration 2u/ul
Source Bacillus species d134
Buffer 33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 55°C. (Incubation at 37°C results in 30% 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.
Storage Buffer 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with BspPI.
Ligation/Recutting AssayAfter 10-fold overdigestion (2u/ug DNA x 5 hours) with BspPI, approximately 80% of the DNA fragments can be ligated in a reaction mixture containing 2040u of T4 DNA ligase/1ug of fragments at a 5'-termini concentration of 0.2uM. More than 50% of these can be recut.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.
Methylation EffectsBspPI does not cut GGm6ATC. Blocked by Dam methylation.
Stability during Prolonged IncubationA minimum of 0.5units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 55°C.
Thermal InactivationEnzyme is inactivated by incubation at 80°C for 20min.
Number of Recognition Sites in DNA
Lambda58
PhiX1740
M13mp18/194
pBR32212
pUC18/1910
pUC5710
pTZ19R/U10
pBluescriptIIKS(-/+)10
pBluescriptIISK(-/+)10
pACYC17713
pACYC1844
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dam-in 1 hour at 55°C in 50ul of assay buffer.


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