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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Bsp68I (NruI)

Bsp68I (NruI)

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Specifications

Catalog #B2935
Sequence5'-T C G^C G A-3'
3'-A G C^G C T-5'
SourceBacillus megaterium RFL68
Concentration10u/ul
Unit DefinitionOne 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.
Diluent Buffer10mM Tris-HCl pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Storage Buffer10mM Tris-HCl pH 7.5, 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol.
R1625 Restriction Enzyme Buffer A, 10X1X buffer formulation - 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
R1625-03 Restriction Enzyme Buffer D, 10X1X buffer formulation - 50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM sodium chloride and 0.1mg/ml BSA.
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 15-fold overdigestion (15u/ug lambda DNA x 1 hour) with Bsp68I (see Star Activity).
Ligation/Recutting Assay
After 10-fold overdigestion (0.6u/ug DNA x 17 hours) with Bsp68I, more than 80% of the digested PhiX174 DNA fragments can be ligated in a reaction mixture containing 20-40u of T4 DNA ligase/1ug of fragments and 10% PEG at a 5'-termini concentration of 0.17uM. More than 90% 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 10 units of Bsp68I for 4 hours.
Blue/White Cloning Assay
pUC57 was digested at a unique site with 10 units of Bsp68I for 1 hour. After religation and transformation < 1% of white colonies were detected.
Star Activity
An excess of Bsp68I (20u/ug DNA x 1 hour) may result in star activity.
Methylation Effect
DamMay overlap - no effect
Dcm, EcoKI, EcoBINever overlaps - no effect
CpGCompletely overlaps - blocked
Incubation Temperature
Incubate at 37°C.
Stability during Prolonged Incubation
A minimum of 0.2 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Digestion of Agarose-embedded DNA
A minimum of 5 units of enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Thermal Inactivation
Inactivated by incubation at 65°C for 20 min.
Number of Recognition Sites in DNA
Lambda5
PhiX1742
M13mp18/190
pBR3221
pUC18/190
pUC571
pTZ19R/U0


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