Unit Definition:One unit is defined as the amount of Bst1107I is required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of recommended reaction buffer.
Form:Supplied as a liquid in 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
Supplied with:R1625: Restriction Enzyme Buffer A, 10X:Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate pH 7.9 at 37°C, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
R1625-03: Restriction Enzyme Buffer D, 10X:Dilute to 1X for use. 1X buffer composition is 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA.
Thermal Inactivation: Enzyme is inactivated by incubation at 80°C for 20 minutes.
Enzyme Properties:Methylation Effects:Dam, Never overlaps - no effectDcm: Never overlaps - no effectCpG: May overlap - cleavage impairedEcoKI: Never overlaps - no effectEcoBI: Never overlaps - no effect
Stability during Prolonged Incubation: A minimum of 0.1units of Bst1107I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Digestion of Agarose-embedded DNA:A minimum of 5 units is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Number of Recognition Sites in DNA:Lambda: 3PhiX174: 0M13mp18/19: 0pBR322: 1pUC18/19: 0pUC57: 0pTZ19R/U: 0
Overdigestion Assay:No detectable change in the specific fragmentation pattern is observed after a 160-fold overdigestion with Bst1107I (10U/ug lambda DNA x 16 hours).
Ligation and Recleavage (L/R) Assay:The ligation and recleavage assay was replaced with LO test after validating experiments showed LO test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.
Labeled Oligonucleotide (LO) Assay:No detectable degradation of single-stranded or double- stranded labeled oligonucleotides occurred during incubation with 10 units of Bst1107I for 4 hours.
Blue/White Cloning Assay:The B/W assay was replaced with L0 test after validating experiments showed L0 test ability to detect nuclease and phophatase activities with sensitivity that equals to that of B/W test.
Storage and Stability:May be stored at 4°C. For long-term storage, aliquot and store at -20°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.
