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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Cfr42I (SacII)

Cfr42I (SacII)

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Specifications

5'-C C G C^G G-3'
3'-G G^C G C C-5'
Catalog #C3350
Concentration 10u/ul
Source E.coli that carries the cloned cfr42IR gene from Citrobacter freundii RFL42
Buffer 10mM Tris-HCl (pH 7.5), 10mM MgCl2 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. For longer periods–the Storage Buffer should be used.
Storage Buffer 10mM potassium phosphate (pH 7.4 at 25°C), 100mM NaCl, 1mM EDTA, 7mM 2-mercaptoethanol, 0.2mg/ml BSA and 50% glycerol.
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 240-fold overdigestion (15u/ug lambda DNA x 16 hours) with Cfr42I.
Ligation/Recutting AssayAfter 50-fold overdigestion (3u/ug DNA x 17 hours) with Cfr42I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.05uM. 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.
Blue/White Cloning AssayThe mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.
Methylation EffectBlocked by CG methylation.
Stability during Prolonged IncubationA minimum of 0.1units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-embedded DNAA minimum of 20units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible EndsBsh1285I
Number of Recognition Sites in DNA
Lambda4
PhiX1741
M13mp18/190
pBR3220
pUC18/190
pUC570
pTZ19R/U0
pBluescriptIIKS(-/+)1
pBluescriptIISK(-/+)1
pACYC1771
pACYC1841
NoteCertain sites in lambda and PhiX174 DNAs are difficult to cleave with Cfr42I, as well as with its prototype SacII.
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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