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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BfmI (SfeI)

BfmI (SfeI)

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Specifications

Catalog #B1086
Description5'-C^T R Y A G-3'
3'-G A Y R T^C-5'
SourceBacillus firmus S8–336
Concentration 10u/ul
Unit DefinitionOne 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.
Diluent Buffer10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol
Storage Buffer10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol
Supplied WithR1625: Restriction Enzyme Buffer A, 10X:
Suplied as a liquid in 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
Enzyme Properties
Stability during Prolonged Incubation
A minimum of 1.0unit 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 20 minutes.
Compatible Ends
C^TGCAG–Alw44I
Number of Recognition Sites in DNA
Lambda38
PhiX1746
pBR3224
pUC574
pUC18/194
pTZ19R/U6
M13mp18/197
Quality Control
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with BfmI (see Star Activity).
Ligation/Recutting AssayThe 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) AssayNo 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 6 months 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.


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