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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BshNI (HgiCI)

BshNI (HgiCI)

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Specifications

5'-G^G Py Pu C C-3'
3'-C C Pu Py G^G-5'
Catalog #B2925
Concentration 10u/ul
Source Bacillus sphaericus Tk 45
Buffer 50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM NaCl 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 7.5 at 25°C), 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 480-fold overdigestion (30u/ug lambda DNA x 16 hours) with BshNI.
Ligation/Recutting AssayAfter 50-fold overdigestion (3u/ug DNA x 17 hours) with BshNI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.24uM. More than 95% 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 EffectsGGm5CGCC and the hemi-methylated sequence GGPyPuCm5C are cleaved by BshNI at a significantly slower rate. Cleavage is blocked when both strands are methylated at positions: 5' GGPyPuCm5C3' 3'm5CCPuPyG G5'. Overlapping Dcm or CG methylation may influence DNA cleavage.
Stability during Prolonged IncubationA minimum of 0.3units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-embedded DNAA minimum of 5units of enzyme is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible EndsAcc65I (G^GTACC), Bsp1407I, KasI (G^GCGCC), Pfl23II, TatI
Number of Recognition Sites in DNA
Lambda25
PhiX1743
M13mp18/197
pBR3229
pUC18/194
pUC574
pTZ19R/U4
pBluescriptIIKS(-/+)4
pBluescriptIISK(-/+)4
pACYC1771
pACYC18410
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.


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