Plasma/SerumCirculatingNucleicAcidPurificationMiniKit(SlurryFormat)Norgen"sPlasma/SerumCirculatingNucleicAcidPurificationMiniKitprovidesafast,reliableandsimpleprocedureforisolatingcirculatingnucleicacids(mRNA,miRNA,exosomalRNA,viralRNA,DNAandviralDNA)fromvariousamountsofplasma/serumrangingfrom0.25mLto2mL. PurificationisbasedontheuseofNorgen’sproprietaryresinastheseparationmatrix.ThekitisabletoisolateallsizesofcirculatingRNA,includingmicroRNA,aswellasallsizesofexosomalRNAandcirculatingDNA.Norgen’sPlasma/SerumCirculatingNucleicAcidPurificationMiniKitprovidesanadvantageoverotheravailablekitsinthatitdoesnotrequireextensiontubesforthepurificationoffree-circulatingNAfromlargesamplevolumes.Free-circulatingNAcanbeisolatedfromeitherfreshorfrozensamplesusingthiskit,andthekitallowsfortheconcentrationofNAthatispresentinlowconcentrations(1-100ng/mLinhumanplasma).Typicalyieldsoffree-circulatingNAwillvarydependingontheinputsample,astheamountofNApresentinplasmaandserumwilldependuponthehealthstatusoftheindividualandthelevelofnucleasespresentintheblood.ThiskitissuitableforthepurificationofnucleicacidsfromserumorplasmapreparedfrombloodcollectedoneitherEDTAorcitrate.PlasmasamplespreparedfrombloodcollectedonheparinshouldnotbeusedasheparincansignificantlyinterferewithmanydownstreamapplicationssuchasRT-PCR.Free-circulatingnucleicacidsinplasmaandserumhavethepotentialtoprovidebioMarkersforcertaincancersanddiseasestates,andincludetumor-specificextracellularnucleicacidfragmentsandfetalDNAinmaternalblood.Free-circulatingDNAisusuallypresentasshortfragmentsoflessthan1000bp,whereasfree-circulatingRNAand exosomal RNAinplasmaandserumareusuallypresentasshortfragmentsoflessthan1000nt.Inaddition,plasmaorserumalsocontainsfree-circulating miRNA thatareassmallas21nt.FurThermore,circulatingviralDNAorviralRNAmaybepresentintheplasmaortheserumofviral-infectedpatients.Norgen’sPlasma/SerumCirculatingNucleicAcidPurificationMiniKitprovidesanefficientmethodforthepurificationofallsizesofthesefragmentedfree-circulatingnucleicacidfromhumanplasmaorserum.Figure1.IsolationandDetectionofCirculatingRNAfromDifferentPlasmaVolumes.Norgen"sPlasma/SerumCirculatingNucleicAcidPurificationMiniKit(SlurryFormat)wasusedtoisolatecirculatingRNAfrom0.25mL,0.5mL,1mLand2mLplasma.ThreemicrolitresofthepurifiedRNAwasthenusedasthetemplateinRT-qPCRreactionstodetectthehuman5Sgene.The5Shousekeepinggenewasdetectedfromallplasmasamplevolumesused.Theamplificationofthe5SrRNAshowedanincreasingamountofRNAwithincreasingthesampleinputvolume.ThisisrepresentedbythedecreaseoftheCtvaluewithincreasingthesampleinputvolume.]">Plasma/SerumCirculatingNucleicAcidPurificationMiniKit(SlurryFormat)Figure1Figure2.EffectiveIsolationofPlasmaCirculatingRNAfromDifferentVolumes.Norgen"sPlasma/SerumCirculatingNucleicAcidPurificationMiniKit(SlurryFormat)wasusedtoisolatecirculatingRNAfrom0.25mL,0.5mL,1mLand2mLplasma.ThreemicrolitresofthepurifiedRNAwasthenusedasthetemplateinRT-qPCRreactionstodetectthehuman5Sgene.The5Shousekeepinggenewasdetectedfromallplasmasamplevolumesused.Theamplificationofthe5SrRNAshowedanincreasingamountofRNAwithincreasingthesampleinputvolume.ThisisrepresentedbythedecreaseoftheCtvaluewithincreasingthesampleinputvolume.]">Plasma/SerumCirculatingNucleicAcidPurificationMiniKit(SlurryFormat)Figure2Figure3.IsolationandDetectionofCirculatingDNAfromDifferentPlasmaVolumes.Norgen"sPlasma/SerumCirculatingNucleicAcidPurificationMiniKit(SlurryFormat)wasusedtoisolatecirculatingDNAfrom0.25mL,0.5mL,1mLand2mLplasma.ThreemicrolitresofthepurifiedDNAwasthenusedasthetemplateinqPCRreactionstodetectthehuman5SrRNAgene.The5Shousekeepinggenewasdetectedfromallplasmasamplevolumesused.Theamplificationofthe5SrRNAgeneshowedanincreasingamountofDNAwithincreasingthesampleinputvolume.ThisisrepresentedbythedecreaseoftheCtvaluewithincreasingthesampleinputvolume.]">Plasma/SerumCirculatingNucleicAcidPurificationMiniKit(SlurryFormat)Figure3