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Pvu I
产品性质
| biological source【生物来源】 | bacterial (Proteus vulgaris) |
| Quality Level【质量水平】 | 100 |
| form【形式】 | solution |
| packaging【包装】 | pkg of 100 U (10650137001 [5 U/μl]) pkg of 500 U (10650129001 [5 U/μl]) |
| manufacturer/tradename | Roche |
| parameter【参数】 | 37 ℃ optimum reaction temp. |
| shipped in【运输】 | dry ice |
| storage temp.【储存温度】 | −20℃ |
基本信息
| Specificity【特异性】 | 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 ℃ for 15 minutes. |
| Quality【质量】 | 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℃ 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【DNA图谱分析】 | Number of cleavage sites on different DNAs
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| Unit Definition【单位定义】 | One Unit is the enzyme activity that completely cleaves 1 μg DNA in one hour at +37 ℃ in a total volume of 25 μl (1x) SuRE/Cut Buffer H. |
| Analysis Note【分析说明】 | 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 ℃), 50 mM KCl, 1.5 mM MgCl2, 200 μM dNTPs, 2.5 U Taq DNA polymerase. The mix was subjected to 25 amplification cycles. 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
Incubation temperature +37℃ Unit definition One Unit is the enzyme activity that completely cleaves 1μg λDNA in 1 hour at +37℃ 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℃. 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℃ in 66mM Tris-HCl, 5mM MgCl2, 5mM Dithiothreitol, 1mM ATP, pH 7.5 (at +20℃) 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. |
| Other Notes【其他说明】 | 仅用于生命科学研究。不可用于诊断。 |
| Components【组分】 | 组份不可单独销售 Enzyme Solution SuRE/Cut Buffer H 10x concentrated |
安全信息
| Storage Class Code【储存分类代码】 | 12 - Non Combustible Liquids |
| WGK | WGK 1 |
