Technical Data
SmiI (SwaI)
Molecular Biology Storage: -20CShipping: Blue Ice
5'-A T T T^A A A T-3'
3'-T A A A^T T T A-5'

Streptococcus milleri S


Unit Definition:
One unit is defined as the amount of SmiI required to digest 1ug of Ad2 DNA-SspI fragments in 1 hour at 30C in 50ul of assay buffer.

Storage Buffer:
10mM Tris-HCl (pH 7.4, 25C) 200mM NaCl, 1mM DTT, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.

Supplied With:
R1625 Restriction Enzyme Buffer A: 10X: 10mM Tris-HCl (pH 7.4, 25C) 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 Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. 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:
30C. Incubation at 37C results in 70% activity.

Methylation Effects:
Dam, Dcm, CpG, EcoKI: never overlaps; no effect.
EcoBI: may 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 30C

Thermal Inactivation:
SmiI is inactivated by incubation at 65C 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:
Lambda: 0
PhiX174: 0
pBR322: 0
pUC57: 0
pUC18/19: 0
pTZ19R/U: 0
M13mp18/19: 1
Ad2: 1
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.