Evaluation of Real-Time PCR Efficiency by the Use of Two Strategies: Standard Curve and Linear Regression

[摘要] BackgroundandObjective:ThedetectionofnucleicacidsusingReal-TimeCPRhasmanyapplication.QuantitativeanalysisofmRNAusingReal-TimePCRbyRelativeandAbsolutemethods.Widelyusedinbiologicalstudies.ThepurposeofthisstudywastocomparetheRelativeandcalculatedPCREfficiencybystandardcurveandLinRegPCRmethods.SubjectandMethods:AftersamplingandextractionofRNAandcDNAsynthesis,thequantitativeRT-PCRwasperformed,thenthePCREfficiencywascalculatedbyUsingthetwomethodsofstandardcurveandLinRegPCR.Attheendtheresultsofthetwomethodswerecomparedandanalyzed.Results:TheefficiencyofPCRfortheGAPDH,TGF-βandIL-10geneswiththestandardcurvemethodwere1.99,1.81and1.87,respectively.ThePCRefficiencyofthesethreegeneswere1.98,1.82and1.82byusingLinRegPCRsoftwaremethod,respectively.Conclusion:TheanalysisofthedataobtainedfromPCRproliferationofcDNAofthethreegenes,GAPDH,TGFβandIL-10showednostatisticaldifferencebetweenstandardcurveandLineRegPCRmethods.Therefore,itisrecommendedtousetheLinRegPCRsoftwaremethodinsteadoftheexpensivestandardcurvemethod.

[学科分类] 基础医学