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Specifications

5'-C^C N N G G-3'
3'-G G N N C^C-5'
Catalog #	B2905
Source	Bacillus stearothermophilus RFL1434
Concentration	10u/ul
Unit Definition	One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 55°C in 50ul of assay buffer.
Dilution Buffer	10mM Tris-HCl, pH 7.4 at RT, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
Storage Buffer	10mM Tris-HCl, pH 7.5 at RT, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
Directions For Use	Incubate at 55°C. (Incubation at 37°C results in 10% activity.)
Supplied with	R1625: Restriction Enzyme Buffer A, 10X. Suitable for use in Double Digest assays.
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 at least 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.
Enzyme Properties
Thermal Inactivation	Enzyme is inactivated by incubation at 80°C for 20min.
Stability during Prolonged Incubation	A minimum of 0.2units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 55°C
Number of Recognition Sites in DNA
Lambda	105
PhiX174	6
M13mp18/19	9
pBR322	8
pUC18/19	5
pUC57	4
pTZ19R/U	5
Methylation Effects
DAM	Never overlaps - no effect
Dcm	May overlap - no effect
CpG	May overlap - no effect
EcoKI	Never overlaps - no effect
EcoBI	Never overlaps - no effect
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 BseDI.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with BseDI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 1uM. 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 restriction endonuclease for 4 hours.