Pvu I

半岛bd体育手机客户端 说明

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.

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图谱分析

Number of cleavage sites on different DNAs

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

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 BufferH.

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 inboldis recommended for optimal activity

• A: 50-75%
• B: 75-100%
• H: 100%
• L: 25-50%
• M: 50-75%

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. coliC 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 MgCl ₂, 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.

其他说明

仅用于生命科学研究。不可用于诊断。

半岛bd体育手机客户端 性质

生物来源	bacterial (Proteus vulgaris)
质量水平	100
形式	solution
包装	pkg of 100 U (10650137001 [5 U/μl]) pkg of 500 U (10650129001 [5 U/μl])
manufacturer/tradename	Roche
参数	37 ℃ optimum reaction temp.
运输	dry ice
储存温度	?20℃

组份列表

组份不可单独销售

货号	组份
	Enzyme Solution
	SuRE/Cut Buffer H 10x concentrated

安全信息

储存分类代码	12 - Non Combustible Liquids
WGK	WGK 1
闪点(F)	does not flash
闪点(C)	does not flash