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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BspLI (NlaIV)

BspLI (NlaIV)

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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 RJ3212
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 AssayNo detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BspLI.
Ligation/Recutting AssayAfter 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) 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 EffectsBspLI cleaves the hemi-methylated sequence GGNNCm5C. Cleavage is blocked when both strands are methylated at positions: 5 GGNNCm5C3' 3'm5CCNNG 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 DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20min.
Number of Recognition Sites in DNA
Lambda82
PhiX1746
M13mp18/1918
pBR32224
pUC18/1911
pUC5712
pTZ19R/U13
pBluescriptIIKS(-/+)15
pBluescriptIISK(-/+)15
pACYC17710
pACYC18423
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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