Singlecellwholegenomeamplification(WGA)withlessbiasmakingthiskitideallysuitedfornextgenerationsequencingapplications.

  • Highsensitivity:Typicalyieldisgreaterthan5µgwhenstartingwithasinglemammaliancell.
  • Reducedamplificationbiasandnoprimerartefacts:Primer-freeamplificationmethodutilizesacombinationofTthPrimPolPrimase(tosynthesizeprimers)andPhi29DNApolymerase.
  • Wellsuitedfornextgenerationsequencing:TestedinIlluminaandIonTorrentNGSworkflows.
  • Easyandreliable
  • InsensitivetoexternalDNAcontamination

  • HowdoesTruePrime™Technologywork?
  • Wholegenomeamplificationfrom5cells
  • Absenceofprimerartefacts
  • InsensitivitytoexternalDNAcontaminations
  • Excellentgenomecoverage

HowdoesTruePrimetechnologywork?

Single Cell Whole Genome Amplification How It Works

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Wholegenomeamplificationfrom5cells

Whole Genome Amplification from 5 cells

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Absenceofprimerartefacts

TruePRime Single Cell Whole Genome Amplification has absence of artefacts

1pgofhumangenomicDNA(~1/6ofthecontentofonehuman/mammaliancell)hasbeenamplifiedusingeitherTruePrime™(TthPrimPol-basedMDA)orrandomprimedMDAreactions.Randomprimedreactionscontain20%ofsequencesthatcannotbemappedtoanyorganisminsequencedatabases.

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InsensitivitytoexternalDNAcontaminations

Duetothehighsensitivityofsinglecellwholegenomeamplificationtechniquesthereisaconsiderabledangerofcontaminatingreactionswithnon-targetderivedDNA.Wethereforerecommendtotakeextremecarewhensettingupsinglecellamplificationreactionsaslaidoutinthehandbook.However,TruePrime™usershaveauniqueadvantagehere:TruePrime™doesnotacceptnon-denaturedDNAstrandsastemplateasreADIlyasrandomprimedMDAreactions.Therefore,contaminationscominginfromtheenvironmentetc.afterthedenaturationstepwillnothaveastronginfluenceonthecompositionofyourreactionoutput.WehaveshownthisbehaviourinanexperimentwherewesimulateanexternalDNAcontaminationbydeliberatelyaddinginyeastDNAtoadenaturedhumanDNAtemplate.YeastDNAisinathousand-foldexcessoverthetargetDNA(1pghumangenomicDNA):

TruePrime Single Cell WGA Insensitive to External DNA Contamination

WhereaswithTruePrime™thevastmajorityofthesequencesobtainedaretarget-derived,randomprimedMDAshowsamajorityofcontaminant-derivedsequences.

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Excellentgenomecoverage

TruePrime Coverage Yeast Chromosome 7

WehaveamplifiedandsequencedtheyeastgenomeusingTruePrime™andIlluminatechnology.Using2.5millionreadpairswecover99.4%ofthetotalyeastgenome.Shownisthecoveragewithnarrowwidthofthedistributioncurve.

Genome coverage of Yeast

Acoveragegraphofyeastchromosome7exemplifiesthemoreevencoverageobtainedbyTruePrime™MDA(middlebluelinerepresentsmeancoverage).

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ORDERINFORMATION

EachTruePrime™SingleCellWGAKitversion2.0contains:BufferL1,DTT,BufferN,ReactionBuffer,dNTPs,Water,Enzyme1,Enzyme2,Control1,andControl2.ThecompletemanualisavailableundertheManualtab.LucigenisanauthorizeddistributorofSygnisproductsintheUS.

TruePrime™SingleCellWGAversion2.0kitisintendedformolecularBIOLOGyuseonlyandinvitrouseonly.Thisproductisnotintendedfordiagnosis,preventionortreatmentofadiseaseinhumanbeingsoranimals.

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