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Technical Data |
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B2905 |
BseDI (SecI) |
300u |
| Molecular Biology | Storage: -20°CShipping: Blue Ice |
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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 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. |
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