Description:
miRNAsareknowntoinvolveinmultipleBIOLOGicalfunctionsandabnormalexpressionisassociatedtohumandiseasessuchascancers.TheyrepresentapromisingnewclassofcancerbioMarkers.AnumberofmultiplexmethodssuchasmicroarrayormicrobeadsassaysarecommonlyusedforprofilingtheexpressionofmultiplemiRNAssimultaneously.HoweverthesemethodsneedtheconversionofthemiRNAintoaCDNAprobeduringanalysis.SignosishasdevelopedasimplemiRNAdirecthybridizationplatearraymethod,inwhichtotalRNAdoesnotneedanypre-treatmentbeforehybridization.Itisassimpleasdirecthybridizationanddetection.Moreimportantly,total48miRNAscanbequantitativelyanalyzedandcomparedsimultaneouslywiththesampleassay.ApplicableGrid:
ListofApplicablemiRNAs
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | let-7b-5p | miR-17-5p | miR-29a-3p | miR-132-3p | miR-182-5p | miR-206 | let-7b-5p | miR-17-5p | miR-29a-3p | miR-132-3p | miR-182-5p | miR-206 |
| B | miR-1 | miR-18a-5p | miR-34a-5p | miR-133 | miR-192-5p | miR-210-3p | miR-1 | miR-18a-5p | miR-34a-5p | miR-133 | miR-192-5p | miR-210-3p |
| C | miR-7-5p | miR-19a-3p | miR-92-3p | miR-143-3p | miR-194-5p | miR-214-3p | miR-7-5p | miR-19a-3p | miR-92-3p | miR-143-3p | miR-194-5p | miR-214-3p |
| D | miR-9-5p | miR-19b-3p | miR-101a | miR-145-5p | miR-196a-5p | miR-222-3p | miR-9-5p | miR-19b-3p | miR-101a | miR-145-5p | miR-196a-5p | miR-222-3p |
| E | miR-10b-5p | miR-20a-5p | miR-106a-5p | miR-146a-5p | miR-199a-5p | miR-223-3p | miR-10b-5p | miR-20a-5p | miR-106a-5p | miR-146a-5p | miR-199a-5p | miR-223-3p |
| F | miR-15a-5p | miR-21-5p | miR-107 | miR-148a-3p | miR-200 | miR-342-3p | miR-15a-5p | miR-21-5p | miR-107 | miR-148a-3p | miR-200 | miR-342-3p |
| G | miR-15b-5p | miR-24-3p | miR-122a-5p | miR-155-5p | miR-204-5p | miR-372-3p | miR-15b-5p | miR-24-3p | miR-122a-5p | miR-155-5p | miR-204-5p | miR-372-3p |
| H | miR-16-1 | miR-26a-5p | miR-125a-5p | miR-181a-5p | miR-205-5p | U6 | miR-16-1 | miR-26a-5p | miR-125a-5p | miR-181a-5p | miR-205-5p | U6 |
Principle:
Signosis’miRNAdirecthybridizationplatearrayisasimpletwo-stepassay;platehybridizationandstreptavidin-HRPdetection.Theplateispre-coatedwithanoligomix,includingapairofuniqueoligosthathybridizeside-by-sidetoaspecifictargetmiRNAandauniversalcaptureoligoandabiotin-labeledoligo.Intheassay,totalRNAisdirectlyutilizedforhybridization.WhenthetargetmiRNAexistsinRNA,itactsasabridgetobringthebiotin-labeledoligotothecaptureoligo,whichcanbedetectedthroughstreptavidin-HRPconjugateandachemiluminescentsubstrate.IfthereexistsnospecificmiRNA,thebiotin-labeledprobewillbewashedaway,leADIngtonodetection.Intheplatearray,48wellsarecoatedwithdifferentoligomixesfordifferentmiRNAs.Asingle96-wellplateallowsquantitativemeasurementandcomparisonof48miRNAsbetweentwosamples.U6RNAisusedfornormalization.
Data:
miRNAdirecthybridizationplatearrayIusedforanalyzingtheexpressionof48miRNAsinhumanskeletalmuscleandkidneytotalRNAs.Asindicatedinthefigure,miR-1andmiRNA133arespecificallyexpressedinskeletalmuscle,whichagreeswithpublishedstudies.
