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Figure1.RNAGelResultsshowing28S:18Sratios.(Imagekindlyprovidedbytebu‐biolaboratory'sRNAextractionandGenomicProfilingServicedivision).
Figure2.ImageJquantificationof28S:18Sratio.Firstmeasurethebackground(left),thenmeasurethe28Sband(middle),thenmeasurethe18Sband(right).Subtractthebackgroundfromthe28Sand18Svaluesandcalculatetheratio.
Figure3.ImageJProfilePlotassessingRNAquality.UsingthisRNAgelimagefoundontheinternet,theImageJ(Analyze‐ProfilePlotFunction)wasusedtodrawahistogram,showingalow28S:18Sratioandsignificantsmearingbelowthe18Sband,indicatingpoorRNAquality.
Figure4.SampleAgilentBioanalyzerResults.Usingamicrofluidicsapproach,theAgilentBioanalyzerdeterminestherelativeabundancesof28Sand18SRNAaswellasthedegreeofdegradation.ThesoftwarecreatesdigitalplotsthatresembletraditionalRNAgels(left)andcalculatestheRNAIntegrityNumber(RIN)basedontheplottedhistogram(right).
Figure5.Meltingcurves.Acceptablemeltingcurvesarethoseshowingameltingcurve(blue)distinctfromthemeltingcurveyougetwiththenegativecontrol(yellow).Agoodprimerpairwillgiveaspecificpeak(left)whileunacceptableprimerpairswillyieldproductswithlesspuremeltingcurves(middleandright).
Figure6.AmplificationCurves.AcceptableamplificationcurvesareC‐shaped(left).cDNAthatistooconcentratedmayyieldanS‐shapedcurve(right),howeverfurtherdilutionofthesampleincreaseCtvaluestothemeasurablerangeof20‐30.
Figure7.qPCRStandardCurves.AcceptablestandardcurveswillhaveanR2near1.0andanEfficiencyof100%orless.
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