Description
5'-A C T G G N^-3' 3'-T G A C^C N-5'
Source
Bacillus species N
Unit Definition
One unit is defined as the amount of BseNI required to digest 1ug of lambda DNA in 1 hour at 65°C in 50ul of assay buffer.
Diluent Buffer
For short term storage (3-4 weeks)- dilute with 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Storage Buffer
For long term storage: 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Supplied With
R1625-01: Restriction Enzyme Buffer B, 10X Suplied as a liquid in 10mM Tris-HCl pH 7.5, 10mM magnesium chloride, 0.1mg/ml BSA.
R1625: 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.
Methylation Effects
Dam: never overlaps - no effect Dcm: never overlaps - no effect CpG: never overlaps - no effect EcoKl: may overlap - effect not determined EcoBl: may overlap - effect not determined
Stability during Prolonged Incubation
A minimum of 0.1units of BseNI is required for complete digestion of 1ug of DNA in 16 hours at 65°C.
Thermal Inactivation
BseNI is inactivated by incubation at 80°C for 20min.
Number of Recognition Sites in DNA
Lambda: 110 M13mp18/19: 18 pBR322: 19 pUC18/19, pUC57: 11 pTZ19R/U: 12 ϕX174: 9
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 BseNI.
Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with BseNI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 1.0uM. 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 10 units of BseNI 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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