BshTI (AgeI)

Artikelnummer: USB-B2930
Artikelname: BshTI (AgeI)
Artikelnummer: USB-B2930
Hersteller Artikelnummer: B2930
Alternativnummer: USB-B2930-200,USB-B2930-1000
Hersteller: US Biological
Kategorie: Molekularbiologie
5-AC C G G T-3 3-T G G C CA-5 Concentration: 10u/ul Source: Bacillus sphaericus Jo 22-024 Buffer: 50mM Tris-HCl pH 7.5, 10mM MgCl2, 100mM NaCl and 0.1mg/ml BSA. Incubate at 37C Diluent Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Storage Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 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, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. R1625-03: Restriction Enzyme Buffer D, 10X: Dilute to 1X for 100% BshTI digestion. 1X buffer composition is 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA. Incubation Temperature: 37C Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with BshTI (see Star Activity). Ligation/Recleavage (L/R) Assay: The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 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 a single-stranded or double-stranded labeled oligonucleotide was observed after incubation with 10units of BshTI for 4 hours. Blue/White Cloning Assay: TheB/W assay was replaced with L0 test after validating experiments showed L0 test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test. Thermal Inactivation: BshTI is inactivated by incubation at 80C for 20 minutes. Stability during Prolonged Incubation: A minimum of 0.1units of BshTI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Compatible Ends: Cfr10I, Kpn2I, Cfr9I, Eco88I, BsaWI, SgrAl, Mrel Methylation Effects: Dam and Dcm: never overlaps - no effect. CpG: completely overlaps - blocked EcoKl and EcoBl: may overlap - effect not determined Digestion of Agarose-embedded DNA: A minimum of 20 units of BshTI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA: Lambda: 13 PhiX174: 0 M13mp18/19: 0 pBR322: 0 pUC18/19: 0 pUC57: 0 pTZ19R/U: 0 M13mp18/19: 0 Unit Definition: One unit is defined as the amount of BshTI required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of reaction buffer. Storage and Stability: Aliquot and 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.