Asingle-ordouble-strandedcircularDNAamplificationkitwithlessbiasandmoreuniformamplificationstartingwithlimitingamountsofDNAorbacterialcells.
- Reducedamplificationbiasandnoprimerartefacts:Primer-freerollingcircleamplification(RCA)methodutilizingacombinationofTthPrimPolPrimase(tosynthesizeprimers)andPhi29DNApolymerase.
- Highlysensitive:AmplificationofattogramamountsofpurifiedinputDNA
- Easyhandlingandhighreproducibility
- InsensitivetoexternalDNAcontaminations
TheTruePrime™RCAkitusesanovelmultipledisplacementamplificationmethodbasedonthecombinationoftherecentlydiscoveredDNAprimaseTthPrimPolandthehighlyprocessiveandhigh-fidelityPhi29DNApolymerasetoamplifysingle-ordouble-strandedcircularDNAmoleculesbyrollingcircleamplification(RCA).ThestrongstranddisplacementcapacityofPhi29DNApolymeraseallowsTthPrimPoltogeneratenewprimersonthedisplacedstrandsthatareextendedbyPhi29DNApol,resultinginexponentialisothermalDNAamplification.TheproductofTruePrime™RCAreactionishighmolecularweight,double-strandedconcatemersofthecirculartemplate.TheTruePrime™RCAkiteliminatestheneedforovernightcellcultureandconventionalDNApurification.TypicalDNAyieldsfroma25µLreactionare>3µgfora3hourreactionwhenstartingfrom1ngofinputplasmidDNA(3-10kb).For150-200kbtemplates,yieldsareapproximately2µgper25µLreactionwith10ngDNAanda3hourreaction.
Suitablestartingmaterialsinclude:
- PurifiedDNA:plasmids,cosmids,BACs,M13clones,etc.
- Bacterialcells:coloniespickedfromagarplates,bacterialculturesorglycerolstocks.
- HowdoesTruePrime™Technologywork?
- ExcellentsensitivitywithplasmidsorBACs
- Absenceofbackgroundamplification
- Plasmidamplificationdirectlyfrombacterialcells
- Lowerbackgroundsandbetteryieldsthancompetitor
HowdoesTruePrimetechnologywork?
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ExcellentsensitivityfromplasmidstoBACs
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Absenceofbackgroundamplification
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Plasmidamplificationdirectlyfrombacterialcells
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Lowerbackgroundsandbetteryieldsthancompetitor
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ORDERINFORMATION
EachTruePrime™RCAKitcontains:BufferD,BufferN,ReactionBuffer,dNTPs,Water,Enzyme1andEnzyme2.ThecompletemanualisavailableundertheManualtab.LucigenisanauthorizeddistributorofSygnisproductsintheUS.
TheTruePrime™RCAkitisintendedformolecularBIOLOGyuseonlyandinvitrouseonly.Thisproductisnotintendedfordiagnosis,preventionortreatmentofadiseaseinhumanbeingsoranimals.
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