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PVUI-RO

Roche

Pvu I

from Proteus vulgaris

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About This Item

UNSPSC Code:
12352204

biological source

bacterial (Proteus vulgaris)

Quality Level

form

solution

specific activity

5000 units/mL

packaging

pkg of 100 U (10650137001 [5 U/μl])
pkg of 500 U (10650129001 [5 U/μl])

manufacturer/tradename

Roche

parameter

37 °C optimum reaction temp.

color

colorless

pH

7.5-7.6 (37 °C)

solubility

water: miscible

suitability

suitable for molecular biology

application(s)

life science and biopharma
sample preparation

foreign activity

Non specific endunuclease <10%
Non specific endunuclease, none detected

storage temp.

−20°C

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.

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.

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

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.

Storage and Stability

Do not store below −25°C.

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.
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.

Other Notes

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

Kit Components Only

Product No.
Description

  • Enzyme Solution

  • SuRE/Cut Buffer H 10x concentrated

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Articles

The term “Restriction enzyme” originated from the studies of Enterobacteria phage λ (lambda phage) in the laboratories of Werner Arber and Matthew Meselson.

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Restriction endonucleases in prokaryotes function primarily to protect against foreign genetic material, notably bacteriophage DNA.

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