CpoI (RsrII)
5'-C G↓G W C C G-3' 3'-G C C W G↑G C-5'
Source
Caseobacter polymorphus
Unit Definition
One unit is defined as the amount of Cpol required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of recommended reaction buffer.
Storage Buffer
10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
R1625: Restriction Enzyme Buffer A, 10X
Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
Incubation Temperature
37ºC
Thermal Inactivation
Cpol is inactivated by incubation at 65°C for 20 minutes.
Diluent Buffer
10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Methylation Effects
Dam, Never overlaps - no effect Dcm: Never overlaps - no effect CpG: Completely overlaps - blocked EcoKI: Never overlaps - no effect EcoBI: Never overlaps - no effect
Stability during Prolonged Incubation
A minimum of 0.5units of Cpol is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Compatible Ends
Cfr13I, Eco47I, Psp5II, SanDI
Number of Recognition Sites in DNA
Lambda: 5 PhiX174: 0 M13mp18/19: 0 pBR322: 0 pUC18/19: 0 pUC57: 0 pTZ19R/U: 0
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion with Cpol (10U/ug lambda DNA x 16 hours).
Ligation/Recutting Assay
The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.
Labeled Oligonucleotide (LO) Assay
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of Cpol for 4 hours.
Blue/White Cloning Assay
The B/W assay was replaced with L0 test after validating experiments showed L0 test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test.
Storage and Stability
May be stored at 4°C. For long-term storage, aliquot and store at -20°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.
Source
Caseobacter polymorphus
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.
