Description:
E.coliUvrDisaDNAhelicasethatusestheenergygeneratedbythehydrolysisofnucleosidetriphosphatestobreakthehydrogenbondslinkingthetwostrandstogetherinduplexDNA.Itisactiveat37ºC.
SUPPLIEDWITH:
10XHelicaseBufferM
SuppliedbyUsers:ATP
Source:
PurifiedfromanE.colistrainthatoverexpressesE.coliuvrDgene.
Reactioncondition:
1XHelicaseBufferM:
10mMDithiolthreitol
10mMMagnesiumAcetate
35mMTris-AcetatepH7.5@25°C
3mMATP.
Incubateat37ºC.
Notes:
E.coliUvrDhelicasemayrequireMutLand/orSSBforoptimalperformanceinspecificapplications.
:MutLstimulatesUvrDhelicaseactivitymorethantenfoldbyloADIngitontotheDNAsubstrate(MechanicLE,FrankelBA,MatsonSW(2000)JBiolChem275:38337–38346).
:MutLandSSBarerequiredinHelicase-DependentAmplificationusingE.coliUvrD(MyriamVincent,YanXu,andHuiminKong(2004)EMBOreportsVol.5,795-800).
StorageConditions:
StorageTemperature:
-20ºC
Concentration:
0.5mg/ml
StorageBuffer:
20mMTris-HClpH8.3
200mMNaCl
1.5mMEDTA
25mMß-Mercaptoethanol
50%Glycerol
QualityControl:
QualityAssuranceStatement:
E.coliUvrDHelicaseispurifiedfreeofcontaminatingendonucleasesandexonucleases.EachlotistestedforDNAunwindingactivity.
