5'-C^C C G G G-3' 3'-G G G C C^C-5'
Unlike SmaI, Cfr9I produces DNA fragments with a 4-base 5'-extension. To achieve complete digestion of substrate with Cfr9I, the concentration of DNA should not be less than 50ug/ml in a reaction buffer.
Source: E.coli that carries the cloned cfr9IR gene from Citrobacter freundii RFL9
Buffer: 10mM Tris-HCl (pH 7.2), 5mM MgCl2, 200mM sodium glutamate 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 Tris-HCl (pH 7.5 at 25°C), 250mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Cfr9I.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Cfr9I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.03uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) Assay
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.
Stability during Prolonged Incubation
A minimum of 0.2units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal Inactivation
Enzyme is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-embedded DNA
A minimum of 5units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Stability During Prolonged Incubation
A minimum of 0.2units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37ºC.
Compatible Ends
BshTI, BsaWI, Cfr10I, Eco88I, Kpn2I, NgoMI, SgrAI.
Number of Recognition Sites in DNA
Unit Definition
One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-HindIII fragments in 1 hour at 37°C in 40ul of assay buffer (containing 2ug DNA fragments).
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