PsyI (Tth111I)

Product Overview

• Article Name: PsyI (Tth111I)
• Biozol Catalog Number: USB-P9181-50
• Supplier Catalog Number: P9181-50
• Alternative Catalog Number: USB-P9181-50-1000
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-G A C N N N G T C-3 3-C T G N N N C A G-5 Source: Pseudomonas syringae Lki1-pH124 Concentration: 10u/ul Unit Definition: One unit is defined as the amount of P9181-50 required to digest 1ug of lambda DNA-Smal fragments in 1 hour at 37C in 50ul of assay buffer. Buffer: Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 50mM sodium chloride, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Supplied with R1625-01 Restriction Enzyme Buffer B, 10X: Supplied as a liquid in 10mM Tris-HCl pH 7.5, 10mM MgCl2, 0.1mg/ml BSA. Incubate at 37C Supplied with R1625 Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Thermal Inactivation: PsyI is inactivated by incubation at 80C for 20min. Enzyme Properties: Methylation Effects: Dam, Dcm, EcoKI: Never overlaps - no effect CpG: May overlap - no effect EcoBI: May overlap - effect not determined Stability during Prolonged Incubation: A minimum of 0.1units of PsyI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Digestion of Agarose-embedded DNA: A minimum of 5 units of PsyI is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Number of Recognition Sites in DNA: Lambda: 2 pBR322: 1 PhiX174: pUC57, M13mp18/19, pUC18/19, pTZ19R/U:: 0 Quality Control: Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (1u/ug lambda DNA x 16 hours) with PsyI. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with PsyII, more than 60% of the DNA fragments can be ligated in a reaction mixture containing 20-40u of T4 DNA Ligase/1ug of fragments and 10% PEG at a 5-termini concentration of 0.33µM. 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 10 units of PsyI for 4 hours. Blue/White Cloning Assay: The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10 units of PsyI for 16 hours. After religation and transformation 1% of white colonies were detected. Storage and Stability: May be stored at 4C. For long-term storage, aliquot and store at -20C. 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.