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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Cfr10I (BsrFI)

Cfr10I (BsrFI)

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Specifications

Catalog #C3275
Sequence5'- R ^C C G G Y -3'
3'- Y G G C C^ R -5'
SourceE.coli that carries the cloned cfr10IR gene from Citrobacter freundii RFL10
Concentration 10 units/ul
Unit DefinitionOne unit is defined as the amount of Cfr10I required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
Incubation Temperature 37°C
Diluent Buffer10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Storage BufferSupplied as a liquid in 10mM potassium phosphate (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Supplied withR1625-45: Restriction Enzyme Buffer for Cfr10I, 10X: Supplied as a liquid in 100mM Tris-HCl, pH7.5, 50mM magnesium chloride, 1M sodium chloride, 10mg/ml BSA, 0.2% Triton X-100.
R1625 Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 330mM Tris-acetate, pH 7.9, 100mM magnesium acetate, 660mM potassium acetate, 1mg/ml BSA.
Enzyme PropertiesStar Activity:
An excess of Cfr10I (10u/ug DNA x 1 hour) or low salt concentration may result in star activity.
Compatible EndsBshTI, BsaWI, Cfr9I, Eco88I, Kpn2I, MreI, NgoMIV, SgrAI.
Methylation EffectsDam, Dcm: Never overlaps - No effect.
CpG: Completely overlaps - Blocked.
EcoKI, EcoBI: May overlap - No effect.
Stability during Prolonged IncubationA minimum of 0.1units of Cfr10I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Digestion of Agarose-embedded DNAA minimum of 5units of Cfr10I is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Number of Recognition Sites in DNALambda: 61
PhiX174: 0
M13mp18/19, pUC18/19, pUC57: 1
pTZ19R/U: 2
pBR322: 7
Note: For cleavage with Cfr10I, at least two copies of its recognition sequence are required.
Thermal InactivationCfr10I is not inactivated by incubation at 80°C for 20min (see Protocol for Inactivation Procedure).
Quality Control
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 5-fold overdigestion (5u/ug lambda DNA x 1 hour) with Cfr10I (see Star Activity).
Ligation/Recutting Assay
After 5-fold overdigestion (5u/ug DNA x 1 hour) with Cfr10I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.5uM. 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 Cfr10I for 4 hours.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.


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