riboprobe in vitro transcription systems protocol

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1、tm016.1210:EIVD_TM.qxd12/1/20109:27AMPage1TechnicalManualRiboprobe®invitroTranscriptionSystemsINSTRUCTIONSFORUSEOFPRODUCTSP1420,P1430,P1440,P1450ANDP1460.PRINTEDINUSA.Revised12/10Part#TM016tm016.1210:EIVD_TM.qxd12/1/20109:27AMPage1Riboprobe®invitroTranscriptionSystemsAll

2、technicalliteratureisavailableontheInternetatwww.promega.com/tbsPleasevisitthewebsitetoverifythatyouareusingthemostcurrentversionofthisTechnicalManual.PleasecontactPromegaTechnicalServicesifyouhavequestionsonuseofthissystem.E-mailtechserv@promega.com.1.Description.......

3、...................................................................................................12.ProductComponents.........................................................................................33.GeneralConsiderations.......................................

4、.............................................4A.PropertiesofPromegaVectorsSuitableforinvitroTranscription..............4B.ApplicationsofPromegaVectors......................................................................6C.GeneralCloningTechniques........................

5、......................................................64.RNATranscriptioninvitro.............................................................................7A.DNATemplatePreparation................................................................................7B.Synthe

6、sisofHigh-Specific-ActivityRadiolabeledRNAProbes...................8C.DeterminingPercentIncorporationandProbeSpecificActivity...............10D.RemovaloftheDNATemplateFollowingTranscription............................13E.RemovalofUnincorporatedNucleotides................

7、.......................................14F.SynthesisofLargeAmountsofRNA..............................................................14G.CappingRNAforinvitroTranslation............................................................155.Troubleshooting........................

8、.......................................................................166.References..................