产品资料

Signosis/Cancer miRNA Array/AP-0003/1 Ea

Description:

MiRNAshavebeenwidelyknowntofunctionaseitheroncogenesortumorsuppressorsundercertainconditions.ThedysregulatedmiRNAshavebeenshowntoaffectthehallmarksofcancer,includingsustainingproliferativesignaling,eludinggrowthsuppressors,resistingcelldeath,andinducingangiogenesis.AnincreasingnumberofstudieshaveidentifiedmiRNAsaspotentialbioMarkersforhumancancerdiagnosis,prognosisandtherapeutictargets.SignosishasdevelopedCancermiRNAArrayincluding132cancer-relatedmiRNAsforcancerresearch.

ApplicableGrid:

ListofApplicablemiRNAs

Let-7a-5p	Let-7b-5p	Let-7c-5p	Let-7d-5p	Let-7e-5p	Let-7f-5p	Let-7g-5p	Let-7i-5p	miR-1	miR-7
miR-9-5p	miR-10a-5p	miR-15a-5p	miR-15b-5p	miR-16-5p	miR-17-5p	miR-18b-5p	miR-18b	miR-19a-3p	miR-19b-3p
miR-20a-5p	miR-21-5p	miR-25-3p	miR-28-5p	miR-34a-5p	miR-99a-5p	miR-122a-5p	miR-124a-3p	miR-125a-5p	miR-125b-5p
miR-126-3p	miR-131	miR-133a-3p	miR-133b	miR-143-3p	miR-145-5p	miR-146a-5p	miR-146b-5p	miR-148a-3p	miR-155-5p
miR-181a-5p	miR-181b-5p	miR-181c-5p	miR-182-5p	miR-192-5p	miR-194-5p	miR-195-5p	miR-199a-5p	miR-199b-5p	miR-199a*-3p
miR-200a-3p	miR-200c-3p	miR-204-5p	miR-206	miR-216-5p	miR-223-3p	miR-224-5p	miR-342-3p	mir-376c-3p	miR-375
miR-9-1-5p	miR-10b-5p	miR-17-3p	miR-22-3p	miR-23a-3p	miR-24-3p	miR-26a-5p	miR-26b-5p	miR-27a-3p	miR-27b-3p
miR-29a-3p	miR-29b-3p	miR-29c-3p	miR-30a-3p	miR-30a-5p	miR-30b-5p	miR-30c-5p	miR-92-3p	miR-92b-3p	miR-93-5p
miR-95-3p	miR-101-1-3p	miR-103-3p	miR-106a-5p	miR-106b-5p	miR-107	miR-128a-3p	miR-128b-3p	miR-132-3p	miR-134-5p
miR-135b-5p	miR-136-5p	miR-137	miR-140-5p	miR-141-3p	miR-142-3p	miR-149-5p	miR-150-5p	miR-151-3p	miR-153-3p
miR-154-5p	miR-181d-5p	miR-183-5p	miR-185-5p	miR-186-5p	miR-188-5p	miR-190-5p	miR-191-5p	miR-196a-5p	miR-196b-5p
miR-197-3p	miR-198	miR-200b-3p	miR-202-5p	miR-203a	miR-205-5p	miR-210-3p	miR-214-3p	miR-215-5p	miR-218-5p
miR-219	miR-221-3p	miR-222-3p	miR-296-5p	miR-372-3p	miR-373-5p	miR-488-5p	miR-100-5p	miR-127	miR-142-5p
miR-31-5p	miR-181a-3p	RNU48

Principle:

miRNAisdifferentfromlargemessengerRNAinthreeaspects;(1)miRNAsaresmallsizemoleculeswithquiteabigdifferenceinabundance,(2)maturemiRNAsco-existwiththeirprecursorpre-miRNAandpri-miRNA,whichonlydifferinlength,and(3)manymiRNAsareverycloselyrelatedinsequences,suchasisoforms,onlydifferingbyoneorafewnucleotides.Therefore,theconventionalmircoarraytechnologiescannotdirectlybeappliedtoanalyzingthesemolecules.AnumberofmiRNAmicroarrayproductsarecommerciallyavailable,buttheyareeithertediousinrequiringpre-isolationofmicroRNA,lackthediscriminativepowertodifferentiatebetweenisoforms,orarenotsensitiveenoughtomonitorlowabundantmiRNAs.

Inourarrayassay,eachmiRNAmoleculeistargetedbytwooligosthathybridizewiththetargetmiRNAtoformaRNA/DNAduplex.Whenthesequencesareperfectlymatched,theoligosarealignedwiththemiRNAandthejointcanbeligatedbyDNAligase(figure1).AsinglenucleotidedifferenceamongmiRNAswillblockeitherthehybridizationortheligation;Thus,miRNAisoformscanbedifferentiated.DuetothesmallsizeofmiRNA,thehybridmightnotbestable;Therefore,weintroducethestackingsequences.Byextendingthesetwooligosalongwiththeircomplementaryoligos,thestABIlityisincreased.Oncethepairofoligosisligated,theligatedmoleculesaresubjectedtolinearamplificationviaT7transcriptionintoRNAinthepresenceofbiotin-UTP,whichareusedasprobesforarrayhybridization.Todifferentiateeachisoform,weassigneduniquetagsequencestotheligationoligos,sothatsinglenucleotidedifferencesareconvertedintouniquetagsequences.Inthisway,eachisoformcanbeeasilydistinguishedbyarrayhybridization.

Theassayprocedureincludesthreesteps:(1)mixthemiRNAwithprovidedoligostoformmiRNA/oligohybrids;(2)selectthehybridsandremovefreeoligos,andligatemiRNA-directedpairingofoligostobecomeasingleDNA;and(3)amplifytheligatedDNAwithT7transcription.

Data:

HumanmiRNAarrayIIIanalysisofmiRNAexpressioninHeGP2,HEK293,andHeLacells.

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Signosis/Cancer miRNA Array/AP-0003/1 Ea

Description:

ApplicableGrid:

Principle:

Data: