|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-G^G N C C-3'
3'-C C N G^G-5'
Restriction Enzyme Buffer A, 10X (R1625): Supplied to simplify buffer selection for double digests.
Number of Recognition Sites in DNA:
Methylation Effects: Cfr13I does not cut GGNCm5C. Blocked by overlapping Dcm and CG methylation.
Stability during Prolonged Incubation: A minimum of 0.3units 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.
Compatible Ends: CpoI, Eco47I, Psp5II, SanDI: G^G(A/T)CC
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Cfr13I.
Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Cfr13I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.3uM. 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.
Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 6-12months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Citrobacter freundii RFL13
Unit Definition: One 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.
Storage Buffer (Form): Supplied as a liquid in 10mM potassium phosphate, pH 7.5 at 25°C, 100mM potassium chloride, 1mM EDTA, 7mM 2-mercaptoethanol, 0.2mg/ml BSA, 50% glycerol.
Diluent Buffer: For short term storage (3-4weeks), use Restriction Enzyme Diluent Buffer, R1625-05: 10mM Tris-HCl (pH 7.4 at 25°C, 100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. For long term storage, use the Storage Buffer should be used.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.