rna gel results showing 28s:18s ratios. (image kindly provided by tebu … · kindly provided by...
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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](https://reader031.vdocuments.mx/reader031/viewer/2022020217/5e435158f943b960f57efcdd/html5/thumbnails/1.jpg)
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](https://reader031.vdocuments.mx/reader031/viewer/2022020217/5e435158f943b960f57efcdd/html5/thumbnails/2.jpg)
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](https://reader031.vdocuments.mx/reader031/viewer/2022020217/5e435158f943b960f57efcdd/html5/thumbnails/3.jpg)
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](https://reader031.vdocuments.mx/reader031/viewer/2022020217/5e435158f943b960f57efcdd/html5/thumbnails/4.jpg)
Figure7.qPCRStandardCurves.AcceptablestandardcurveswillhaveanR2near1.0andanEfficiencyof100%orless.