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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » SmiI (SwaI)

SmiI (SwaI)

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Specifications

5'-A T T T^A A A T-3'
3'-T A A A^T T T A-5'
Catalog #S1014-59
SourceStreptococcus milleri S
Concentration10units/ul
Unit DefinitionOne unit is defined as the amount of SmiI required to digest 1ug of Ad2 DNA-SspI fragments in 1 hour at 30°C in 50ul of assay buffer.
Storage Buffer10mM Tris-HCl (pH 7.4, 25°C) 200mM NaCl, 1mM DTT, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
Supplied WithR1625 Restriction Enzyme Buffer A: 10X: 10mM Tris-HCl (pH 7.4, 25°C) 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. For longer periods, the Storage Buffer should be used.
R1625-03 Restriction Enzyme Buffer D 10X: 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA.
Storage and StabilityMay be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Overdigestion Assay
No detectable change in the specific fragmentation pattern was observed after 160-fold overdigestion (10u/ug Ad2 DNA x 16 hours) with S1014-59.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with S1014-59, more than 80% of the DNA fragments can be ligated in a reaction mixture containing 2040u of T4 DNA Ligase/1ug of fragments and 10% PEG at a 5'-termini concentration of 0.06uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) Assay
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide occured during incubation with 10 units of S1014-59 for 4 hours.
Blue/White Cloning Assay
The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10 units of SmiI for 16 hours. After religation and transformation, the background level of white colonies was <1%.
Incubation Temperature
30ºC. Incubation at 37ºC results in 70% activity.
Methylation Effects
Dam, Dcm, CpG, EcoKINever overlaps; no effect.
EcoBIMay overlap; effect not determined
Stability during Prolonged Incubation
A minimum of 0.1 unit of SmiI is required for complete digestion of 1ug of Ad2 DNA in 16 hours at 30°C
Thermal Inactivation
SmiI is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-embedded DNA
A minimum of 10 units of SmiI is required for digestion of 1ug of agarose-embedded Ad2 DNA in 16 hours.
Number of Recognition Sites in DNA
Lambda0
PhiX1740
pBR3220
pUC570
pUC18/190
pTZ19R/U0
M13mp18/191
Ad21


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