Description:

MicroRNAs(miRNAs)aresmallnoncodingRNAsthatcontrolgeneexpressionattheposttranscriptionallevelthroughselectivelybindingtocomplementarymessengerRNAsequences. Approximately30%ofmammaliangenesareregulatedbythesesmallRNAmolecules,which affectavarietyof BIOLOGicalfunctions,includingdevelopment,celldifferentiation,proliferation,apoptosis,andmaintenanceofstemnessandimprinting. 
NorthernblottinghasbeenthemostcommonlyusedmethodforanalyzingindividualmiRNAs,althoughrecentlyanumberofnewapproacheshavebeendeveloped, suchas therelatedreal-timePCRandinvaderassays. LowsensitivityandpoorthroughputarethemainissuesofNorthernblotanalysis. Toresolvethisproblem, Signosishasdevelopedthe miRNAplateassaykits.Thekitsarehighlysensitive,andthebiochemicalconversionofmiRNAmoleculesintoCDNAsisnotneeded.Theprocedureoftheassayisassimpleasincubationandwash.

Principle:

IntheproprietarymiRNAplateassay,onemiRNAmoleculeisflankedbyacaptureoligoandabiotinateddetectionoligothroughtwobridgesoligos.OneofthebridgeoligosispartiallyhybridizedwiththemiRNAmoleculeandthecaptureoligoandanotherwiththemiRNAandthedetectionoligo.Thehybridisimmobilizedontoplatethroughhybridizationwithanimmobilizedoligoanddetectedbyastreptavidin-HRPconjugateandchemiluminecscentsubstrate.ThishybridstructureissensitivetothesequenceofthemiRNAmolecule.OnenucleotidedifferencewillpreventtheformationofthehybridandthereforemiRNAisoformcanbedifferentiated,whichnormallyishardtotacklewithNorthernblot.Inaddition,thesensitivityoftheassayishigherthanmiRNANorthernblotassay.

Data:

miRNAplateassay.BothmiR-133andmiR-1arepreferentiallyexpressedincardiacandskeletalmusclesandplayessentialrolesindifferentiation,proliferationandapoptosisofcardiaccells.DysregulatedmiRNAexpressionhasbeencorrelatedtodiseasedheartsinhumanpatientsandtherefore,theyarewithgreatpotentialsasdiagnosticMarkersandtherapeutictargetsforcardiovasculardisease.(A)SchematicediagramofmiRNAplateassay.(B)ExpressionofmiR-133andmiR-1wasanalyzedwith5ugoftotalRNApreparedfromhumansckeletalmuscle,heart,andliverthroughNorthernblot(top)andmiRNAplatassay(bottom).(C)DetectionofmiR-133expressionwascomparedwithNorthernblot(top)andmiRNAplateassay(bottom).
 

Signosis 是一家专业的生物分析供应商，致力于基于微孔板分析技术的生命科学研究产品的开发、生产和销售。Signosis 成立于2007年，坐落于世界研发中心--美国硅谷的中心地带。公司专注于细胞因子、转录因子、miRNA及疾病标志物等重要调控因子和标志物的研究，基于自有专利技术，开发完成400多种先进而独特的用于发现、筛选和分析的生命科学研究产品。Signosis 产品销售遍布北美、欧洲、亚洲及中东等主要国家和地区，客户涵盖美国国立卫生研究院、哈佛大学、耶鲁大学等著名研究机构以及众多制药公司