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Protocolofthereal-timePCRfordetectinggeneticallymodifiedplantsandtheirderivedproductsT9JMfMfSiSflF*&*a3.1AmpEraseUNC891uracilN-glycosylase.3.2C,@:C:cycle,t:threshold,S+E~WXBrl%%B~Bl&%~Brl~f$BifEM!!5BrlBCIS~.3.31PC:internalpositivecontrol,&#me3.4NTC:notemplatecontrol,9fiMR.3.5PCR:polymerasechainreaction,m$$PCR.3.6DNAdeoxyribonucleicacid,li#N@#@&.3.7dNTPadeoxyribonucleosidetriphosphate,li#N&%=@@3.8dATP:deoxyadenosinetriphosphate,jl#NJBW%E%O&.3.9dCTP8deoxycytidinetriphosphate,B!Nl@BZ@@*3.10dG?'P,deoxyguanosinetriphosphate,ENgl$%Z@&.3.11dUTPrdeoxyuridinetriphosphate,j@,#91%jZ%OBgp.3.12bpgbasepair,MBN.3.13~a~maDNAfiCe@.3.14Tris:tris(hydroxymethy1)aminomethane,.Z(E2:!Ffr)aS:!Sm3.15TEIT~~~-C~,EDTA~#~~3.16Lectintl#B#€#.3.17ZEIN:#@B#EB.3.18P~t~~Pcase:phosphoenilpyruvatecarboxylase,~@~~~~~~lf13PB~~.3.19tRNALeu:&#W8#HB*3.2018srRNA:&@&@18s@@#RNA&B$.3.21CaMV35S:35Spromoterfromcauliflowermosaicvirus,%ElTZ#%ZiV%3i$H35SG3Fe3.22NOS:terminatorofnopalinesynthasegene,MlB@igiff&$tsl)SEl%kF.3.23FMV35SZ35Spromoterfromamodifiedfigwortmosaicvirus(cau1imovirusgroup),Z#ZUj-J$j3335S!aS!JT.3.24NPTII:neomycin-3'-phosphotransferasegene,%~~-3'-@@$$ffl%&H.t3.25BAR:phosphinothricinacetyltransferasegenefromBacillusamyloliquefaciens,S/E8Tf-BacillusamyloliquefaciensfTRZB%$$%hBA2&H03.26PAT*phosphinothricinacetyltransferasegene,*Tf#2m$$bAgm.3.27GOX!glyphosateoxidoreductasegene,fgB.$I&BEmSHa3.28CryIA(b):asyntheticgeneencodesthefirst648aminoacids,insecticidal-activeftruncatedproductidenticaltothatofcrylA(6)geneofBacillusthuringiensissubsp,~Z&~j$f-~~&~~0cryIA(b)SEl.3.29Cry3A:B.thuringiensissubsp,Tenebrionis(B.t.t.strainBI256-82B&eE0,i55;##?@&$t~#$~~#~~&(Coloradopotatobeetlelarvae).3.30EPSPS:5-enolpyruvylshikimate-3-phosphatesynthasegene,5-%j3@-3-@@ff&@&&m.&QB#bIlIflPaq%HEl&%%W%%PCRZB&B~8B'IBfi?34&#$fihk88SN/T1193HdB.#%B#bUIf&q%&H&%%tl$%%PCRZ#&BEMSN/T1194WeSH3%&Tje6BBhS6.1Biia%W%%SBPCR@*,SB&PCREG#%cfrBUA%%gBt+'JH%%@%BR%W%WB+PCRB@,igt%%a6S@&M%@@&SfiSB%fiH3%.PCRF#W&BRA-~~I@HHW~~A-+M%BB~J%~A#~%SH%-~&B@,~~%~I]~3z-+fie%%&&ma-+@x%*BmeS#%sW,fi&sm~%BtJ%%B%@%xsm%&lPCRF#W,TaqAH5'M-3'%9bMABB#S~AM~RW~@fi&%%S~~@K%*6~%R.M~%~PfiS%W~#C9J%%B%~EPSF#--%DNA4S,%G-+%*%F3&,%%?%%#+HRR%PCRf@%&%*Hi6.6.2zswa6.2.1%W%%BSPCRR.6.2.2&8+1#6.6.2.3m3i$xl@iO6.2.4%J@;U1.6.2.5&BQtPQ%6!X.6.2.6&~lCS&Rd.~#l,f?$/l\~B~~$L,Mini+A&~b#l.6.2.7Rl@SPC,lCSiB***%o6.2.8%*#,Zl%*%$6.2.9E&EPlj#.26.2.10~f~##~O.5pL,2pL,10pL,20pL,100pL,2OOpL,1OOOpL.6.2.11*fi.rjc:PCREGB.6.3flSM~PiA*~afi.~;as%$le~alfi~~aYfiafpl.6.3.1X@M;r)tt&R&GB/T6682$Jfi--@;rltMB;gP.6.3.2PCRS*#k.6.3.3#k@(M&lr1.6.3.4dNTPs(dATP.dUTP.dCTP,dGTP).6.3.5UNGE(Uraci1N-glycosylase).6.3.6TaqE.6.3.75I@jinSHB@W#AUIP&$JRHHR%%NR%PCRUiNfiM5I@jinSHR:P!!RS1SHR*PCRSPPJRRllSI#@4R+t!F9JY1aHZEINLectinPE3-PEPcasetRNALeu18srRNACaMV35SNOS:FMV35SNPTllBarPATCOXJI1Q%PJ5'-tgaacccatgcatgcagt-3'5'-ggcaagaccattggtga-3'5'-cctcctcgggaaagttacaa-3'5'-gggcatagaaggtgaagtt-3'5'-ccagttcttggagccgcttga-3'5'-aagggccagtccaaatgcaga-3'5'-~~aaatcggtngacgctacg-3'5'-ttccattgagtctctgcacct-3'5'-cctgagaaacggctaccat-3'5'-cgtgtcaggattgggtaat-3'5'-~gacagt~gtcccaaaga-3'5'-aagacgtggttggaacgtcttc-3'5'-atcgttcaaacatttggca-3'5'-attgcgggactctaatc'ata:3'5'-aagacatccaccgaagactta-3'5'-aggacagctcttttccacgtt-3'5'-aggatctcgtcgtgacccat-3'5'-gcacgagg~gcggtca-3'5'-acaagcacggtcaacttcr3'-5'-actcggccgtc~a~tcgtr3'5'-gtcgacatgtctccggagag-3'5'-gcaaccaaccaagggtatr3'5'-gtcttcgtgttgctggaaccgtt-3'5'-gaactggcaggagcgagagct-3'#H%N5'-tggcgtgtccgtccctgatge3'5'-ccctcgtctcttggtcgcgccctct-3'5'-caggtcgctatgcgactgcggagaca-3'5'-gcaatcctgagcca~tce3'5'-tgcgcgcctgctgccttcct-3'5'-tggacccccacccacgaggagcatr3'5'-catcgcaagaccggc~cagg-3',5'-tggtccccacaagccagctgctcga-3'5'-cacccagccggccacagtcgat-3'5'-ccgagccgcaggaaccgc(lggag-3'5'-tggccgcggtttgtgatatcgttaa-3'5'-tgctcacgttctctacactcgcgctcg3'3883181h*BC#$I:P&(Alsa)RrzB#$I:Pdb(AlaBB)#%Sfa#$I:P&(AlsEi)tta(APMSBB)&#Pa(APMirrRU)%SEIRP,3i*,LISf.#%.W#*B#bnIP&%sakI.3i*nB%Sf.#%.9**BC#$IPB%SB#%b-9#*.3#;dfi%I#&(Sf)B#bnI:Pdassa#%Sf.#;b6.~#IB~#~~IP&esaMsSf.r*aft$IP~%aHRl.3i*.L%SfBB~~I:P&%SH#%Sf.3*B#$IP~6.4B@%S6.4.1MBMR,R~~HjiaPhZ$MMiftB6.4.1.1MtPHRUU%A~~E%SB~@(@AG.~%.~%~)~.-~@PB.B%.M~)#$ULP:~~DNA%&iE06.4.1.2RtPMR%H$~R~SB@~JM&@#$ULP:~~DNA#%PCREGH&&6i%k%H%3%%MSEl@9JMW&.6.4.1.3PBNHsfi?q-@@@DNAN8EM@@2aNM(H7kRB&8)9=@PCRBGH2EIN%(U&RBDNA&&).6.4.2BWmPCRE&#SXNR%PCRtSi&#%%$t2,@+#&B.t8rr=+f%?Be$~#EWGf!l!#apDNA%31E%+bABQ%+96S%MTXSI9$a&GG@#S]%tBYa2awsxPCREB~UWSICry3ACryIA(b)EPSPS6.4.3SWB*PCRE&#&~libf~~~fPCRHBG@#!i%:37C,5min;Bg#k95C,3minr95T.15s,60C91minq404.B,&mB#~#%BaE*#E&#B*3%B@e4B+t#PJ5'-acctatgctcaagctgccaacaccc-3'5'-ccgccgcgagctgaccctgaccgtg-3'5'-ccgcgtgccgatggcctccgca-3'41%BP15'-tccggttacgaggttctt-3'5'-ccatagatttgagcgtcctta-3'5'-cgcgactggatcaggtaca-3'5'-tggggaacaggctcacgat-3'5'-ccgacgccgatcaccta-3'5'-gatgccgggcgtgttgag-3'is#&%IoxPCRtii&S@RRkR(MgC&dNTPs(*dUTP)UNC894128J(l#)5l#J(-F#)SJRQtTuq89DNA#@(20ng-30ng/pL)+E*BSHSB%SB%##BXlnIPka$$SqXXBXMIPAi$$SEiklulk#&IPd%1x2.5mmol/L0.2mmol/L0.075U0.2pmoI/~0.2prnoI/~0.1pmol/L2.5U5PL50p~B:S'+DNA#S%~~~#&~@~~~RIP.P~NB~~UIBBS~M~R#S%.6.4.4BBYilJla&B#PCRgGB@&@9&SM3i,G%@~&MH6Sm