Technical Data
BseDI (SecI)
Molecular Biology Storage: -20CShipping: Blue Ice
5'-C^C N N G G-3'
3'-G G N N C^C-5'

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 55C 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: Supplied as a liquid in 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 55C. (Incubation at 37C 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 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for 6 months after receipt at -20C. 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 80C 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 55C

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
The Ligation/Recutting assay was replaced with LO test after validating experiements showed LO test ability to trace nuclease and phophatase activities with sensitivity that is higher than L/R by a factor of 10.

Labeled Oligonucleotide (LO) Assay:
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of BseDI for 4 hours.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.