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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BseSI

BseSI

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Specifications

G A
5'-G T G C C^C-3'
3'-C^A C G G G-5'
C T
Catalog #B2918
Concentration 10u/ul
Source Bacillus stearothermophilus Jo 10–553
Buffer 10mM Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA. Incubate at 55°C. (Incubation at 37°C results in less than 20% 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, or Storage Buffer.
Storage Buffer 10mM Tris-HCl (pH 7.5 at 25°C), 100mM 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 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with BseSI (see Star Activity).
Ligation/Recutting AssayAfter 50-fold overdigestion (3u/ug DNA x 17 hours) with BseSI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.1uM. 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.
Star ActivityA large excess of enzyme (10u/ug DNA x 16 hours) may result in star activity.
Methylation EffectSequence GKGCm5AC is cleaved by BseSI at a significantly slower rate. Overlapping EcoKI methylation may influence DNA cleavage.
Stability during Prolonged IncubationA minimum of 0.1units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 55°C.
Thermal InactivationOnly small amounts of enzyme (up to 10units) can be inactivated 80°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 EndsGGGCC^C–ApaI, Eco24I, SduI GTGCA^C–Alw21I, Mph1103I, PstI, SdaI, SduI
Number of Recognition Sites in DNA
Lambda10
PhiX1741
M13mp18/192*
pBR3223
pUC18/193
pUC574
pTZ19R/U2
pBluescriptIIKS(-/+)3
pBluescriptIISK(-/+)3
pACYC1772
pACYC1841
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 55°C in 50ul of assay buffer.
* According to our experimental data, BseSI has only one recognition site in M13mp18/19 DNA at a position 2088.


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