MCLAB/T4 Endonuclease VII/ENDO7-200/10,000 U,MCLab,,MCLab,Molecular Cloning Laboratories,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：MCLAB/T4 Endonuclease VII/ENDO7-200/10,000 U

产品编号：ENDO7-200

市场价：¥12600.00

场地：美国(厂家直采)

产品分类：分子类>其他分子产品>寡聚核酸>

联系QQ：1570468124

电话号码：4000-520-616

邮箱： info@ebiomall.com

美元价：$630.00

品牌： MCLab

公司：MCLAB.Inc

公司分类：

商品介绍

EndonucleaseVIIistheproductofgene49ofbacteriophageT4.Ithasamassof18kDa.T4EndonucleaseVIIinvolvesinDNA-packaging,geneticrecombinationandmismatchrepairinvivo.

Description:
EndonucleaseVIIistheproductofgene49ofbacteriophageT4.Ithasamassof18kDa.T4EndonucleaseVIIinvolvesinDNA-packaging,geneticrecombinationandmismatchrepairinvivo.Ithasalsobeendemonstratedinvitrotoresolvesingle-basemisparings,heteroduplexloopsandbranchedDNAs,suchasfour-wayHollidayjunctionsandthree-wayY-structures.

CRISPR-Cas9 System

Source:
Arecombinant E.coli straincarryingtheclonedT4EndonucleaseVIIgene

UnitDefinition:
Oneunit(0.5ng)oftheenzymeresolves50%of1pmolFAMlabeled28meroligonucleotidesubstrate[1]withintheimmobile4-wayHollidayjunctionstructurein30minutesat37°Cin50mMTris-HCl,pH8.0,10mMMgCl2,10mM2-MEand0.1μg/μlBSA.

SpecificActivity: 2000U/µg

RecommendedStorageCondition: -20°C

ExperimentalData:
T4 Endonuclease VII

Figure1. PerformanceofMCLAB’sT4EndonucleaseVIIanalyzedbycapillaryelectrophoresis.(a)Negativecontrolsampleanalysis,10pmolofFAMlabeled28meroligonucleotidesubstrate.(b)40UofMCLAB’sT4EndonucleaseVIIwasabletofullyresolve10pmolofFAMlabeled28meroligonucleotidesubstratewitha4-wayHollidayjunctionstructure(30minutesat37°Cin50mMTris-HCl,pH8.0,10mMMgCl2,10mM2-MEand0.1μg/μlBSA).
T4 Endonuclease VII
Figure2. MCLAB’sT4EndonucleaseVIIandT7EndonucleaseI(fromothersupplier)activitycomparison. (a)10U(275fmol)ofMCLABT4’sEndonucleaseVIIcanresolve64%of10pmolof4-wayjunctionsubstrate(enzymetosubstratemolarratio1:70)in30minutesat37°Cin50mMTris-HCl,pH8.0,10mMMgCl2,10mM2-MEand0.1μg/μlBSA.(b)40U(368fmol)ofT7EndonucleaseI(supplierN)onlyresolves43%of10pmolof4-wayjunctionsubstrate(enzymetosubstratemolarratio1:50)in30minutesat37°Cin50mMNaCl,10mMTris-HCl,pH7.9,10mMMgCl2,1mMDTT.ComparedtoT7EndonucleaseI(supplierN),MCLAB’sT4EndonucleaseVIIresolves4-wayHollidayJunctionsatahigheryieldin30minuteswithlessenzyme.
T4 Endonuclease VII
T4EndonucleaseVIIProtocol

For4-wayjunctionsubstrate:

1. Preparethefollowingreaction:

Component	Volume
Water	16µl
10xEndoVIIreactionbuffer	2µl
10µM4-wayJunction(FAMlabeled)	1µl
EndoVII(diluteifnecessary)	1µl
Total	20µl

2. Mixwellandincubatethereaction@37°Cfor30min.

3. Use1µlreactiontoanalyzethecleavedfragmentsonCapillaryelectrophoresis.

Forsinglebasemismatchsubstrate:

1. Useabout200-400ngDNAfragmentthatcontainssinglebasemismatchforeachreaction:

Component	Volume
10xEndoVIIbuffer	1µl
Substrate	200-400ng
EndoVIIenzyme(diluteifnecessary)	1µl
Water	Tototalvolumeof10µl
Total	10µl

2. Mixwellandincubatethereaction@37°Cfor30min.

3. Runa2%agarosegelandcheckforcleavedbands.

Table.ComparisonofT4EndonucleaseVIIandT7EndonucleaseI

	T4EndonucleaseVII	T7EndonucleaseI
ProteinMass	18KDa	60kDa
Function	Resolvase	Resolvase
Application	Enzymaticmutationdetection ResolvebranchedDNA Detectorcleaveheteroduplexes	Enzymaticmutationdetection ResolvebranchedDNA Detectorcleaveheteroduplexes
Source	BacteriophageT4	BacteriophageT7
ProteinDesign	T4EndonucleaseVII	Afusionofmaltosebindingprotein(MBP)andT7EndonucleaseI
Activity (Testedwith4-wayHollidayjunctionsin30minutes.)	High* 10unitsoftheenzyme(278fmolprotein,molarratio1:70(Enzyme:Substrate))resolve64%of4-wayJunctions.	Low* 40unitsoftheenzyme(368fmolprotein,molarratio1:50(Enzyme:Substrate))resolve43%of4-wayJunctions.
Specificity	High*(singleresolvedpeakshownonCE)	Low*(multipleresolvedpeaksshownonCE)

*SeeFigure2forT4endonucleaseVIIandT7endonucleaseIactivitycomparisonresult.

Suppliedin:
10mMTris-HCl
50mMKCl
1mMDTT
0.1mMEDTA
50%Glycerol
pH7.4@25°C

SuppliedWith:
10xT4EndonucleaseVIIReactionBuffer:
500mMTris-HCl
100mMMgCl2
100mM2-mercaptoethanol
1mg/mlBSA
pH8.0@25°C

Reference:
1.Golz,S.,Birkenbihl,R.P.,andKemper,B.(1995),DNAResearch2,277-284.

品牌介绍

关于我们

Established in 1998, Molecular Cloning Laboratories (MCLAB) is a leader in providing genomic research consumables and services to the Life Sciences community. MCLAB provides cost-effective consumables and reagents, focusing on first generation as well as next-generation sequencing and DNA fragment analysis. We manufacture a large selection of molecular biology related products, including antibodies, biochemical reagents, cloning kits, enzymes, PCR kits, protein gels & solutions and more. With a team of over twenty experienced scientists and over six decades of experience, MCLAB is committed to offering first-rate services to researchers. We specialize in the field of genomics, with experts in DNA sequencing, fermentation, fragment analysis, molecular cloning, over-expression, protein purification, and protein synthesis. MCLAB has offices in China and distributors all around the world, including Australia, Austria, Belgium, Canada, Czech Republic, Denmark, Germany, Hungary, Greece, India, Israel, Poland, New Zealand, Turkey, United Kingdom and Vietnam.

网站http://www.mclab.com

行业生物技术

规模11-50 人

总部South San Francisco， CA

类型私人持股

创立1998

领域Next-generation Sequencing， DNA fragment analysis， Antibodies， Biochemical Reagents， Cloning Kits， Enzymes， PCR Kits， Protein Gels & Solutions， DNA Sequencing， Fermentation， Molecular Cloning， Protein Purification， Protein Synthesis