PVUI-RO

Pvu I

from Proteus vulgaris

• Codice UNSPSC: 12352204

Proprietà

• Origine biologica: bacterial (Proteus vulgaris)
• Livello qualitativo: 100
• Forma fisica: solution
• Attività specifica: 5000 units/mL
• Confezionamento: pkg of 100 U (10650137001 [5 U/μl]); pkg of 500 U (10650129001 [5 U/μl])
• Produttore/marchio commerciale: Roche
• Parametri: 37 °C optimum reaction temp.
• Colore: colorless
• pH: 7.5-7.6 (37 °C)
• Solubilità: water: miscible
• Compatibilità: suitable for molecular biology
• applicazioni: life science and biopharma; sample preparation
• Attività estranea: Non specific endunuclease <10%; Non specific endunuclease, none detected
• Temperatura di conservazione: −20°C

Descrizione

Specificità

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.

Qualità

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.

Profilo DNA

Number of cleavage sites on different DNAs

• λ: 3
• φX174: 0
• Ad2: 7
• M13mp7: 1
• pBR322: 1
• pBR328: 1
• pUC18: 2
• SV40: 0

Definizione di unità

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.

Stoccaggio e stabilità

Do not store below −25°C.

Risultati analitici

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.

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.

Altre note

For life science research only. Not for use in diagnostic procedures.

Solo come componenti del kit

• Enzyme Solution
• SuRE/Cut Buffer H 10x concentrated

Informazioni sulla sicurezza

• Codice della classe di stoccaggio: 12 - Non Combustible Liquids
• Classe di pericolosità dell'acqua (WGK): WGK 1
• Punto d’infiammabilità (°F): does not flash
• Punto d’infiammabilità (°C): does not flash