I.U.B.:3.1.21.1
Bovinepancreaticdeoxyribonucleaseisanendonucleasewhichsplitsphosphodiesterlinkages,preferentiallyadjacenttoapyrimidinenucleotideyieldingpolynucleotideswithfreehydroxylgroupatthe3"positionandphosphategroupatthe5"position.Theaveragechainlengthofalimitdigestisatetranucleotide.
Uses:WorthingtonoffersDNaseatdifferentlevelsofpurityfordifferentapplications.ProductCodes:DPRFandDPRFSarebothespeciallydesignedforMolecularBIOLOGyapplicationsandcontainthelowestlevelsofRNaseandproteaseactivity.TheyarebothsuitableforuseintechniquesrequiringdigestionofDNAintherecoveryofintactRNAorwheretheintegrityofstructuralproteinsorenzymesmustbemaintained.Applicationshaveincludednicktranslation,DNAmapping,isolationofnuclearRNAandprotein,plasmidconstruction,andRNApolymerasesynthesisofRNAprobesandRT-PCR.DNaseisalsousedintissuecultureworktodigestDNAfromdamagedcellstherebyreducingviscosity,andremovingmembraneboundDNAfragments.WorthingtonCodes:DPandDCLSaresuitablefortheseapplications.
Stability/Storage:Whenproperlystored,allgradesofWorthingtondeoxyribonucleasearestablefor2-3years.RecommendedstoragetemperatureforallgradesofWorthingtonDNasesis2-8°C.ProductcodeDPRFSmaybestoredat-20°C.Forlongtermstorageinsolution,ProductCodesDandDPFFmaybedissolvedin5mMacetate,1mMcalcium,pH4.5andstoredinsingleusealiquotsat-20°Cor-70°Cforuptooneyear.Onlyfreezeandthawonce;thawedaliquotsarestablerefrigeratedatleastseveralweeks.Additionof50%glycerolwillmaintainaliquidstateat-20°Cwithoutaffectingstabilityandmaterialin50%glycerolcanberemovedandreturnedto-20°Crepeatedly.DPRFisunusuallystableduetotheabsenceofprotease.ForlongtermstorageofDPRFafterreconstitution,usewateroranybufferpH4.0to9.0exceptphosphate;avoidcalciumchelators;add50%glycerolforstorageasliquidat-20°C;aliquotinsingleusecontainers;onlyfreezeandthawonce;thawedaliquotsarestablerefrigeratedatleastseveralweeks.
UnitDefinition:1Unitcausesanincreaseinabsorbanceat260nmof0.001perminutepermlat25°CwhenactinguponhighlypolymerizedDNAatpH5.0.Note:Kunitzunitsasreportedbyothersupplierscanbe2to4timeshigherthanKunitzunitsasmeasuredatWorthington.AsmeasuredatWorthington,oneKunitzunitdigests1µgoflamBDaDNAin10minutesat37°Cin50mMTris,1mMMg2+,1mMCa2+,pH7.8ina50µlreaction.CorrelationofdigestionunitswithKunitzunitsisdifferentforotherDNAandbuffersystems.
TechnicalNote:ProductCodeDPRF:Eachvialcontainsapproximately2mgglycineand2µmolescalciumper10,000unitsofDNaseI.Dissolvingtheentirevialin5mlprovidestheequivalentofa1mg/mlsolution.
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