|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-C^C N N G G-3'
3'-G G N N C^C-5'
Source: Bacillus stearothermophilus RFL1434
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.)
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.
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:
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
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BseDI.
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.