Pvu I
ROCHE/PVUI-RO - from Proteus vulgaris
Product Type: Chemical
| application(s) | life science and biopharma sample preparation |
| biological source | bacterial (Proteus vulgaris) |
| color | colorless |
| foreign activity | Non specific endunuclease <10% |
| Non specific endunuclease, none detected | |
| form | solution |
| manufacturer/tradename | Roche |
| packaging | pkg of 100 U (10650137001 [5 U/μl]) |
| pkg of 500 U (10650129001 [5 U/μl]) | |
| parameter | 37 °C optimum reaction temp. |
| pH | 7.5-7.6 (37 °C) |
| Quality Level | 100  |
| solubility | water: miscible |
| specific activity | 5000 units/mL |
| storage temp. | −20°C |
| suitability | suitable for molecular biology |
| Analysis Note: | Compatible ends Pvu I generates ends that are compatible with fragments generated by Pac I. Isoschizomers Pvu I is an isoschizomer to BspC I and Xor II. Methylation sensitivity Pvu I cleavage is not inhibited by overlapping dam-methylation at the site indicated (°) on the recognition sequence, but Pvu I fragments of DNA isolated from dam+ strains are not as readily religated as those isolated from dam- strains. Pvu I is inhibited by 5-methylcytosine at the indicated site (°) and by 4-methylcytosine. SuRE/Cut Buffer System The buffer in bold is recommended for optimal activity • A: 50-75% • B: 75-100% • H: 100% • L: 25-50% • M: 50-75% Incubation temperature +37°C Unit definition One Unit is the enzyme activity that completely cleaves 1μg λDNA in 1 hour at +37°C in a total volume of 25μl SuRE/Cut Buffer H. Heat inactivation Pvu I cannot be heat inactivated by incubating it for 15 minutes at +65°C. PFGE tested Pvu I has been tested in Pulsed-Field Gel Electrophoresis (on bacterial chromosomes). For cleavage of genomic DNA (E. coli C 600) embedded in agarose for PFGE analysis, we recommend 10U of enzyme/μg DNA and 4 hour incubation time. Ligation and recutting assay Pvu I fragments obtained by complete digestion of 1 μg pBR322 DNA are ligated with 1U T4 DNA Ligase in a volume of 10μl by incubation for 16hours at +4°C in 66mM Tris-HCl, 5mM MgCl2, 5mM Dithiothreitol, 1mM ATP, pH 7.5 (at +20°C) resulting in >80% recovery of 1μg pBR322 DNA fragments. Subsequent re-cutting with Pvu I yields >95% of the typical pattern of pBR322 DNA × Pvu I fragments. |
| Analysis Note: | Activity in PCR buffer: <5% Relative activity in PCR mix (Taq DNA Polymerase buffer) is less than 5%. The PCR mix contained λDNA, primers, 10 mM Tris-HCl (pH 8.3, 20 °C), 50 mM KCl, 1.5 mM MgCl2, 200 μM dNTPs, 2.5 U Taq DNA polymerase. The mix was subjected to 25 amplification cycles. |
| DNA Profile: | Number of cleavage sites on different DNAs • λ: 3 • φX174: 0 • Ad2: 7 • M13mp7: 1 • pBR322: 1 • pBR328: 1 • pUC18: 2 • SV40: 0 |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Quality: | Absence of nonspecific endonuclease activities 1μg λDNA is incubated for 16hours in 50μl SuRE/Cut Buffer H with an excess of Pvu I. The number of enzyme units which do not change the enzyme-specific pattern is stated in the certificate of analysis. Absence of exonuclease activity Approximately 5μg [3H] labeled calf thymus DNA are incubated with 3μl Pvu I for 4hours at +37°C in a total volume of 100μl 50mM Tris-HCl, 10mM MgCl2, 1mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis. |
| Specificity: | Pvu I recognizes the sequence CG°AT↓ *CG and generates fragments with 3′-cohesive termini. Recognition sites: CG°AT*CG CG°AT*CG Restriction site: CG°AT↓*CG CG°AT↓*CG Heat inactivation: No inactivation of Pvu I after incubation at 65 °C for 15 minutes. |
| Storage and Stability: | Do not store below −25°C. |
| Unit Definition: | One Unit is the enzyme activity that completely cleaves 1 μg DNA in one hour at +37 °C in a total volume of 25 μl (1x) SuRE/Cut Buffer H. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| activity | specific activity: 5000 units/mL |
| Storage Temp. | −20°C |
| UNSPSC | 12352204 |
| Components | Enzyme Solution; SuRE/Cut Buffer H 10x concentrated |