U6_5S_miRNA 引物探针设计

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1、内参1.RNU6HomosapiensRNA,U6smallnuclear1(RNU6-1),smallnuclearRNANR_004394.1ORIGIN1gtgctcgcttcggcagcacatatactaaaattggaacgatacagagaagattagcatggc61ccctgcgcaaggatgacacgcaaattcgtgaagcgttccatattttRNU6-FCTCGCTTCGGCAGCACARNU6-RAACGCTTCACGAATTTGCGT2.5SHomosapiensRNA,5Sr

2、ibosomal2(RNA5S2),ribosomalRNANR_023364.1ORIGIN1gtctacggccataccaccctgaacgcgcccgatctcgtctgatctcggaagctaagcagg61gtcgggcctggttagtacttggatgggagaccgcctgggaataccgggtgctgtaggctt121t5S-FTCTCGTCTGATCTCGGAAGCTA5S-RGCGGTCTCCCATCCAAGTAMiRNA方案1RTprimeradaptorCTCAACTGGTGTC

3、GTGGAGTCGGCAATTCAGTTGAGXXXXXXForwardprimeradaptorACACTCCAGCTGGGXXXXXXXXXXXXXXUniversalReverseprimerTGGTGTCGTGGAGTCG方案2RTprimeradaptorGTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACXXXXXXForwardprimeradaptorGCGGCGGXXXXXXXXXXXXXXUniversalReverseprimerGTGCAGGGTCCGA

4、GG1.hsa-miR-122序列：uggagugugacaaugguguuug长度：222.Hsa-miR-16序列：uagcagcacguaaauauuggcg长度：223.hsa-mir-93序列：caaagugcuguucgugcagguag长度：23方案1设计引物PrimerResult:miRNAmiRNAsequence(5'-3')PrimerPrimerSequence(5'-3')LengthofprimerTmofprimerGCcontentofprimerhsa-mir-122TGGAG

5、TGTGACAATGGTGTTTGRTprimerCTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGCAAACACC440.522727272727ForwardprimerACACTCCAGCTGGGTGGAGTGTGACAATG29900.55hsa-mir-16TAGCAGCACGTAAATATTGGCGRTprimerCTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGCGCCAATA440.522727272727ForwardprimerACACTCCAGCT

6、GGGTAGCAGCACGTAAAT29880.52hsa-mir-93CAAAGTGCTGTTCGTGCAGGTAGRTprimerCTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGCTACCTGC440.545454545455ForwardprimerACACTCCAGCTGGGCAAAGTGCTGTTCGTG30940.57方案2设计引物窗体顶端PrimerResult:miRNAmiRNAsequence(5'-3')PrimerPrimerSequence(5'-3')Lengt

7、hofprimerTmofprimerGCcontentofprimerhsa-mir-122TGGAGTGTGACAATGGTGTTTGRTprimerGTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCAAACA500.54ForwardprimerGCGGCGGTGGAGTGTGACAATG22720.64hsa-mir-16TAGCAGCACGTAAATATTGGCGRTprimerGTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGA

8、CCGCCAA500.58ForwardprimerGCGGCGGTAGCAGCACGTAAAT22700.59hsa-mir-93CAAAGTGCTGTTCGTGCAGGTAGRTprimerGTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCTACCT500.56ForwardprimerGCGGCGGCAAAGTGCTGTTCG