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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Bsp143I (MboI, Sau3AI)

Bsp143I (MboI, Sau3AI)

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Specifications

Catalog #B2950
Description5'-^G A T C-3'
3'-C T A G^-5'
SourceBacillus species RFL143
Concentration 10u/ul
Unit DefinitionOne unit is defined as the amount of Bsp143I required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
Diluent BufferFor short term storage (3-4 weeks)- dilute with 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
Storage BufferFor long term storage: 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM DTT, 0.1mM EDTA, 0.15% Triton X-100, 0.2mg/ml BSA, 50% glycerol.
Supplied WithR1625-35 Restriction Enzyme Buffer for Bsp143I, 10X:
Suplied as a liquid in 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 0.02% Triton X-100, 0.1mg/ml BSA.
R1625 Restriction Enzyme Buffer A, 10X:
Suplied as a liquid in 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Enzyme Properties
Compatible Ends
BamHI, BcII, BgIII, Psul
Methylation Effects
DamCompletely overlaps - no effect
DcmNever overlaps - no effect
CpGMay overlaps - blocked
EcoKlNever overlaps - no effect
EcoBlMay overlap - no effect
Stability During Prolonged Incubation
A minimum of 0.1units of Bsp143I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal Inactivation
Bsp143I is inactivated by incubation at 65°C for 20min.
Number of Recognition Sites in DNA
Lambda116
M13mp18/197
pBR32222
pUC18/19, pUC57, pTZ19R/U15
X1740
NoteDpnl, Bsp1431 and Mbol all recognize the same sequence but have different methylation sensitivites and cleavage sites.
Quality Control
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Bsp143I.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bsp143I, more than 95% of the digested DNA fragments can be ligated at a 5'-termini concentration of 1.2uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) Assay
No detectable degradation of single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of Bsp143I for 4 hours.


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