Technical Data
B1086-05
BfuI (BciVI)
100U
Molecular Biology Storage: -20CShipping: Blue Ice
Sequence:
5'-G T A T C C (N)6^-3'
3'-C A T A G G (N)5^-5'

Source: Bacillus firmus Auk

Concentration: 5u/ul

Unit Definition:
One unit is defined as the amount of BfuI required to digest 1ug of lambda DNA dcm in 1 hour at 37C in 50ul of reaction buffer.

Supplied With:
R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (pH 7.9 at 37C).

R1625-20: Restriction Enzyme Buffer for Bful, 10X: Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25C), 300mMNaCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.

Storage and Dilution Buffer:
10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.

Thermal Inactivation:
Bful is inactivated by incubation at 80C for 20min.
Overdigestion Assay:
No detectable change in the specific fragmentation pattern is observed after 5-fold overdigestion (5u/ug lambda DNA x 1 hour) with BfuI.

Ligation/Recutting Assay:
After 5-fold overdigestion (5u/ug DNA x 1 hour) with BfuI, more than 80% of the DNA fragments can be ligated in a reaction mixture containing 2040u of T4 DNA ligase/1ug of fragments and 10% PEG at a 5'-termini concentration of 0.25uM. More than 90% 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 BfuI for 4 hours.

Star Activity:
A large excess of BfuI (7.5u/ug DNA x 1 hour) may result in star activity.

Stability during Prolonged Incubation:
A minimum of 1.0unit of BfuI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C.

Digestion of Agarose-embedded DNA:
A minimum of 5units of BfuI is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Recommended Protocol for Digestion:
Add:
Nuclease free water: 16ul
R1625-20: 2ul
DNA (0.5-1ug/ul): 1ul
Bful: 0.5-2ul
Mix gently, spin down for a few seconds.
Incubate at 37C for 1-16 hours.

Protocol for Digestion of PCR products directly after amplification:
Add:
PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA)
Nuclease free water: 18ul
R1625-20: 2ul
Bful: 1-2ul
Mix gently, spin down for a few seconds.
Incubate at 37C for 1-16 hours.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.