Psp1406I (AclI)

5'-A A^C G T T-3' \n3'-T T G C^A A-5'\n\nSource: \nPseudomonas species RFL1406\n\nConcentration: 10u/ul\n\nIncubation Temperature: 37 degrees C\n\nUnit Definition: One unit is definied as the amount of Psp1406I required to digest 1 ug of lambda DNA in 1 hour at 37 degrees C in 50ul of assay buffer.\n\nDiluent Buffer: \n10mM Tris-HCl (pH 7.4 at 25 degrees C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.\n\nStorage Buffer: \nSuplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25 degrees C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.\n\nSupplied with: \nR1625: Restriction Enzyme Buffer A, 10X:\nSuplied as a liquid in 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.\n\nEnzyme Properties\nCompatible Ends: \nBsp119I, Bsu15I, Hin1I, Hin6I, HpaII, MspI, NarI, Ssil, TaqI, XmiI\n\nMethylation Effects: \nDam, Dcm: Never overlaps - No effect.\nCpG: Completely overlaps - Blocked.\nEcoKI, EcoBI: May overlap - Effect not determined. \n\nStability during Prolonged Incubation: \nA minimum of 0.5 units of Psp1406I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37 degrees C. \n\nThermal Inactivation: \nPsp1406I is inactivated by incubation at 65 degrees C for 20min. \n\nDigestion of Agarose-embedded DNA: \nA minimum of 10 units of Psp1406I is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. \n\nNumber of Recognition Sites in DNA:\nLambda: 7\nPhiX174: 3\nM13mp18/19: 2\npBR322: 4\npUC18/19: 2\npUC57: 2\npTZ19R/U: 2\npBluescriptIIKS(-/+): 2\npBluescriptIISK(-/+): 2\npACYC177: 2\npACYC184: 2\n\nQuality Control:\nOverdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Psp1406I. \n\nLigation and Recleavage (L/R) Assay: \nThe L/R 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.\n\nLabeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of Psp1406I for 4 hours.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for 6 months at -20 degrees 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.

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