真菌检测鉴定通用引物-Fungal-Primers模板.doc

真菌检测鉴定通用引物-Fungal-PrimersITS1:5’-TGCGG-3’ITS4:5’-GCTTATTGATATGC-3’Tm55℃NS17:CATGTCTAAGTTTAAGCAANS3:S4:TTTAAGNS22:AATTAAGCAGACAAATCACTNS24:TTgTTACgACTTTTALR0R:5’-GCTGAACTTAAGC-3’LR3:5’-CCGTGTTTCAAGACGGGLR3R:5’-plementarytoRLR3R:GTGTTTCAAGAC)LR5:5’-TTAAAAAGCTCGTAGTTGAAC-3’LR7:5’-AAGATCTLR12:5’-GACTTAGAGGCGTTCAGLr0R/LR5:Tm50-52℃NL1:5’-GCATATCAATAAGCGGAGGAAAAGNL1:5′-TGC(alsocalledV9:TGC)NL1:5’-ATGGATC-3NL2:5’-CTCTCTTTTCAAAGTTCTTTTCATCTNL2:5’-AACGGCTTCGACAACAGC-3NL2:5’-CTTGTTCGCTATCGGTCTC(alsoNL2A:5′-CTTGTTCGCTATCGGTCTC)NL2:5’-TACTTGTTCGCTATCGGTCT-3'NL3:5’-GATAGCGAACAAG(alsoNL3A:5’-GATAGCGAACAAG)NL3:5’-GATAGCGAACAAGTANL3:5’-GTT-3NL4:5’-GTGTTTCAAGACGG(similartoRLR3R:5′-GTGTTTCAAGAC)NL4:5’-TAGATACATGGCGCAGTC-3ConservedprimersequencesforPCRamplificationandsequencingfromnuclearribosomalRNA()Vilgalyslab,DukeUniversityOvertheyears,roupsoffungi(primarilyEumycota),,plantsandothereukaryotes;,theseprimersspanmostofthenuclearrDNAcodingregion(seefigures),;variablepositionsareindicatedasfollows:P=A,G/Q=C,T/R=A,T/V=A,C/W=G,"R"representthecodingstrand(sameasRNA).-encodedribosomalRNAgenes(rDNA)offungiexistasamultiple-prisedofhighlysimilarDNAsequences(typicallyfrom8-12kbeach)arrangedinahead-to-(containingtheprimaryrRNAsforasingleribosome),punctuatedbyoneormoreintergenicspacer(IGS)(ycetousyeasts),