MCLAB/T4核酸内切酶VII/ENDO7-100/2500 U-免疫在线蚂蚁淘旗下平台

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商品详细MCLAB/T4核酸内切酶VII/ENDO7-100/2500 U

MCLAB/T4核酸内切酶VII/ENDO7-100/2500 U

商品编号： ENDO7-100

品牌： Molecular Cloning Laboratories

市场价： ¥4200.00

美元价： 2520.00

产地：美国(厂家直采)

公司：

产品分类：寡聚核酸

公司分类： Oligomeric_nuclei_acid

联系Q Q： 3392242852

电话号码： 4000-520-616

电子邮箱： info@ebiomall.com

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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

MCLAB (Molecular Cloning Laboratories) is a leader in DNA sequencingservices and consumables. We are committed to providing the bestservices and products to researchers worldwide. MCLAB's large capacityDNA sequencing services (10,000+ reactions in 24 hours) is well knownfor high quality (up to 1000 bp read length), low cost (as low as$3.50/reaction) and fast turn around time (8~24 hours). Our proprietaryDNA sequencing consumables include NanoPOP (DNA Sequencing and FragmentAnalysis Matrix), NanoPOP Running Buffer, BigDye Dilution Buffer, CARE(Capillary Array Regeneration Excellence) solution, and DNA SizeStandards. In Addition, MCLAB also provides many other services,including gene synthesis, protein expression and purification, and codonmaximization. Other products include PCR, Real-time PCR, frozencompetent yeast, E. coli, as well as novel and traditional enzymes formolecular cloning applications.

公司介绍

MCLAB (Molecular Cloning Laboratories) is a leader in DNA sequencingservices and consumables. We are committed to providing the bestservices and products to researchers worldwide. MCLAB

品牌分类

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