| Catalog # | C3300 |
| Sequence | 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. |
| Enzyme Properties | Number of Recognition Sites in DNA: |
| Lambda: 74 |
| PhiX174: 2 |
| pBR322: 15 |
| pUC57: 8 |
| pUC18/19: 6 |
| pTZ19R/U: 6 |
| M13mp18/19: 4 |
| 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 |
| Quality Control | 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. |
| Source | |
| Citrobacter freundii RFL13 |
|
| Concentration | ~10u/ul |
|
| 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. |
