RatDigestivetissueNorthernblotismadeoftotalRNAextractedfrom11digestivetissuesofSpragueDawleyrat(Panel#4).TotalRNAsamplesaresubjectedtorigorousqualitycontrolprocedures beforefractionationthroughlargedenaturedagarosegels. BlotscanbeusedwitheitherrADIoactiveor nonisotopic-labeledprobesto:
Quicklyassessgeneexpressionacrossrattissues.
Determinesizeandrelativeabundanceoftranscriptsindifferenttissues.
Identifyalternativespliceforms.
Qualitycontrol:
Everystepofpreparationofblots,fromharvestingtissuesandextractionofRNAtotheblotting,iscarefullymonitoredtoensurethesuperiorqualityandperformance.
Blotsaremade oftotalRNAthatistreatedwithRNase-freeDNasetoremoveresidualDNA.ThepurityandintegrityofRNAaretestedbydenaturingagarosegelelectrophoresis.
TheefficiencyoftransferischeckedbystaininggelswithreversIBLestaining.
TheintegrityoftheblottedRNAistestedbybeta-actinspecificprobe.
Description: ThefreshlyisolatedtotalRNAsamples(20ugeach)plusoneRNAMarkerarefractionatedthroughlarge formaldehyde-denaturedagarosegeleletrophoresis,blottedontoapositivelychargednylonmembranes,andimmobilizedbyUVlight.Blotisshippedinasealedbagatroomtemperatureand isgoodfor1yearifproperlystoredat4oCinthesealedbag.