MCLAB/2X Taqman probe based HoTaq qPCR Kit/HTP430RF/1 Ea

ForResearchUseOnly.

Figure:AmplificationofhumanGAPDHgenetargetwith2XHoTaqReal-timePCRKit.Amplificationcurvesareshownfortenfolddilutionsof0.0002pMto20pMofplasmid.Insetshowsthestandardcurvedata.

Description:
Thisisahighperformancereal-timePCRreagent.ItutilizesMCLAB'sproprietaryquantitativePCRtechnology.

Advantages:
2xHoTaqPCRReactionMixproductsaresuperiorinamplifyingdifficulttemplatescomparingwithsimilarproductsfromothersuppliers.

-ThisistheamplificationofGPIIBgene(70%G+C).
-10~10Kcopiesfrom30pghumangenomicDNAhavebeendetected.

Application:
ProbebasedquantitativePCR:includingDNAquantification,2-stepRTPCR,SNPanalysis,etc.

Primerandprobedesign:
1.Toachievethebestperformance,appropriatesoftware,suchasABIPrimerExpress^TM,shouldbeusedtodesignprimerswith50°C~65°Cannealingtemperatureand68°C~70°Cforprobeswith17~30nucleotidesinlength
2.Ampliconsizeshouldbesmall,<150bp
3.Avoidsecondarystructuresinprimersandprobes
4.Avoidmorethan3consecutiveGsinprimersandprobes
5.Primersshouldnothavecomplementary3'-ends
6.17~30nucleotidesinlength

RecommendedReactionConditions::
95°C,10minutes.->(95°C,5seconds.->60°C,30seconds.)for50cycles.

RecommendedStorageCondition:-20°C

Notes:
Toachieveaccuratequantification,itishighlyrecommendedtodoreplicatesandtoreducepipettingerrors.

Reference:
1.Holland,P.M.,Abramson,R.D.,Watson,R.,andGelfand,
2.D.H.1991.ProceedingsoftheNationalAcademyofSciencesUSA88:7276-7280.
3.Livak,K.J.,Flood,S.J.A.,Marmaro,J.,Giusti,W.,andDeetz,K.1995.PCRMethodsandApplications4:357-362.
4.Lee,L.G.,Connell,C.R.,andBloch,W.1993NucleicAcidsResearch21:3761-3766.