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Figure 1. RNA Gel Results showing 28S:18S ratios. (Image kindly provided by tebu‐bio laboratory's RNA extraction and Genomic Profiling Service division). Figure 2. ImageJ quantification of 28S:18S ratio. First measure the background (left), then measure the 28S band (middle), then measure the 18S band (right). Subtract the background from the 28S and 18S values and calculate the ratio.

RNA Gel Results showing 28S:18S ratios. (Image kindly provided by tebu … · kindly provided by tebu‐bio laboratory's RNA extraction and Genomic Profiling Service division). Figure

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Page 1: RNA Gel Results showing 28S:18S ratios. (Image kindly provided by tebu … · kindly provided by tebu‐bio laboratory's RNA extraction and Genomic Profiling Service division). Figure

Figure1.RNAGelResultsshowing28S:18Sratios.(Imagekindlyprovidedbytebu‐biolaboratory'sRNAextractionandGenomicProfilingServicedivision).

Figure2.ImageJquantificationof28S:18Sratio.Firstmeasurethebackground(left),thenmeasurethe28Sband(middle),thenmeasurethe18Sband(right).Subtractthebackgroundfromthe28Sand18Svaluesandcalculatetheratio.

Page 2: RNA Gel Results showing 28S:18S ratios. (Image kindly provided by tebu … · kindly provided by tebu‐bio laboratory's RNA extraction and Genomic Profiling Service division). Figure

Figure3.ImageJProfilePlotassessingRNAquality.UsingthisRNAgelimagefoundontheinternet,theImageJ(Analyze‐ProfilePlotFunction)wasusedtodrawahistogram,showingalow28S:18Sratioandsignificantsmearingbelowthe18Sband,indicatingpoorRNAquality.

Figure4.SampleAgilentBioanalyzerResults.Usingamicrofluidicsapproach,theAgilentBioanalyzerdeterminestherelativeabundancesof28Sand18SRNAaswellasthedegreeofdegradation.ThesoftwarecreatesdigitalplotsthatresembletraditionalRNAgels(left)andcalculatestheRNAIntegrityNumber(RIN)basedontheplottedhistogram(right).

Page 3: RNA Gel Results showing 28S:18S ratios. (Image kindly provided by tebu … · kindly provided by tebu‐bio laboratory's RNA extraction and Genomic Profiling Service division). Figure

Figure5.Meltingcurves.Acceptablemeltingcurvesarethoseshowingameltingcurve(blue)distinctfromthemeltingcurveyougetwiththenegativecontrol(yellow).Agoodprimerpairwillgiveaspecificpeak(left)whileunacceptableprimerpairswillyieldproductswithlesspuremeltingcurves(middleandright).

Figure6.AmplificationCurves.AcceptableamplificationcurvesareC‐shaped(left).cDNAthatistooconcentratedmayyieldanS‐shapedcurve(right),howeverfurtherdilutionofthesampleincreaseCtvaluestothemeasurablerangeof20‐30.

Page 4: RNA Gel Results showing 28S:18S ratios. (Image kindly provided by tebu … · kindly provided by tebu‐bio laboratory's RNA extraction and Genomic Profiling Service division). Figure

Figure7.qPCRStandardCurves.AcceptablestandardcurveswillhaveanR2near1.0andanEfficiencyof100%orless.