Description:

IsoAmpIIIUniversaltHDAkitcanbeusedtoamplifyanddetectshortDNAsequences(70bp-120bp)ataconstanttemperature.Thekitcanbeusedwithavarietyoftemplates,includingmicrobialgenomicDNA,viralDNA,plasmidDNA,andCDNA.AsinglecopyoftargetDNAcanbeamplifiedbytHDAanddetectedbyagarosegelelectrophoresiswhenoptimizedprimersandbufferareused.
IsoAmp®IIIUniversaltHDAkitisbasedonathird-generationtHDAformulaformorerobustandspecificamplificationwithreducednon-specificproductsduetoprimer-dimer.Insomecases,rapidamplificationmaybeachievedbyaddingmoreIsoAmp®IIIEnzymeMixintothereaction(unpublishedobservation).ThevolumeofIsoAmp®IIIEnzymeMixhasbeenreducedto2µLperreaction,whichallowstheusertoaddmoreenzymes(2Xor3X)forpossIBLehigh-speedamplification.IsoAmp®IIIEnzymeMix(Cat#H1020S)canbepurchasedseparately.
IsoAmp®IIIUniversaltHDAkitsupportsallreactionswhicharesupportedbyIsoAmp®IIUniversaltHDAkit.Selectedexamplesareincludedinthismanual,includingend-pointtHDAandreal-timequantitativeHDA(qHDA)inbothone-stepandtwo-stepformats.AControlTemplateandasetofspecificprimersaresuppliedforthepositivecontrolreaction.



Figure2.PositivecontrolassayswithvaryingamountofControlTemplate.
Positivecontrolassayswereconductedfollowingtheone-stepqHDAprotocol.ControlTemplatewasusedat5ng-500fgperassayintriplicate.
*Negativecontrolreactionsmayproduceprimer-dimer,whichcanbedistingushedfromspecificampliconsbygel-analysisorbymelting-curveanalysis.
Applications:
UsedtoamplifyanddetectshortDNAsequences(80-120bp)
-downloadManual
Advantages:
UseofhelicasetoseparateDNAeliminatesneedforThermocycler
Reactionscanbeperformedatconstanttemperature.
Workswithavarietyoftemplates,includingbacterialgenomicDNA,viralDNA,plasmidDNAandcDNA.
AslowasasinglecopyoftargetDNAcanbeamplifiedunderoptimizedconditions.
Canbeusedforreal-timequantitativeor/andreversetranscriptionHDA.
KitComponents:
StorageConditions:
QualityControl:
QualityAssuranceStatement:
KitcomponentswereindividuallytestedforoptimalperformanceintHDA.
Legal:
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