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ProtocolofthequalitativepolymerasechainreactionfordetectinggeneticallymodifiedplantcomponentsinedibleoilT~UXf++8tl&%~~*~SRtsfi5IH~B%*6@HA@RS&1#9HSIHX#t#M,GEJ?%HB&$(&Q%~iBHb@)%Bi5~%&;BHT*ERt%Thitgh@@%*ER%BB~H83HRE3W@Hjh34rX#HS%M*oRE$&H#9H511X#t#t:a%M*3HT*6@oGB/T6682j)~g!&SJ#&~#@~Bfj-&(eqvIS03696:1987)SN/T1193S~@~~!&~~~~~SN/T1194~@W~F&%~BB%@MM#I#r~1#2i%SN/T1195kS+ffSBB%SBPCR&8!!2it&SN/T119633R+ffSB&%Z#PCRB@l3%SN/T1197?&%H+ff&BB%SBPCRBM2i&!i-SN/T1199#j%+%&Hfi&%Z#@B!lPCR3%SN/T1202.$t&+%SB&@&%Z#PCRW!!I3%SN/T1204&$hW#$QIFaa+%SBB%AB$?EfiPCR&B@4$2i%T3U#i~%SHT*h5Ro3.1PCR:polymerasechainreaction,m#&PCR.3.2DNA:deoxyribonucleica~id,NR@@j&@~3.3dNTP:deoxyribonucleosidetriphosphate,ag@%Z@@.3.4dATP:deoxyadeNOSinetriphosphate,%!@.8$%=€#@3.5dCTP:deoxycytidinetriphosphate,$!glfi%3€#@.,3.6dGTP:deoxyguaNOSinetriphosphate,li#RsffZ@@.3.7dUTP:deoxyuridinetriphosphatel~~Jj25jff:€#@3.8bp:basepair,&Sjt;f.3.9Taq88i:NNAB&88.3.10Tris:tris(hydroxymethyI)aminomethane,3(~PS)$ilaSPj$.,3.11TE:Tris-CI.EDTA%#$&.3.12Lectin:B@@%%l.3.13ZEIN:&$QB$%EQ3.14PE3-PEPcase:phosphoenilpyruvateCarboxylase,@@#€!$&fiBA@ftkII.3.15tRNALeu:B@W%#SiH3.16CaMV35s:35spromoterfromcauliflowermdsaicvirus,*$TE##%EWSic5535sG33.3.17NOS:terminatorofnopalinesynthasegene,fdBB#&-$?&Bi&B%It3.3.18FMV35S135Spromoterfromamodifiedfigwortmosaicvirus(caulimovirusgroup),x&EWI%t35SB325;b3.3.19NPTII:neomycin-3'-phosphotransferasegene,%S!#-3'-@BS@BSH.3.20BAR:phosphinothricinacet~ltransferasegenefromBacillusamyloliquefaciens,*@IBacillusamyloliquefaciensTB2i%%@ISH3.21PAT,phosphinothricinacetyltransferasegene,@TM2%S$$i@SB.3.22GOXtglyphosateoxidoreductasegene,#i'HI#$!,kSEi@SE!lai6.1JHB@iC&&H3tBflR%SBr$DNAh3;-@BRaH85SSiH@@EMR%Ac559hNi&B.tftff41C,%S$-&AfgjCLZG(PcR)B*,%R&rMfiISafHDNAER,@BPCRfMBWC,Pl@fB&Ha1PcfiEB$%SSH&3306.2f#W#6.2.1PCR%m9W.6.2.2DNA3WBW.6.2.3B?91#&.6.2.4~SXBIBiWI.6.2.5~@PB5j&W.6.2.6$$i!E~tftEB~b#L,13i%/1\?4!&jL4l,,Mini+hBjL4l,.6.2.7fiB%qpC,@B@rft;b)(Ro6.2.8%&#,A%tk%#6.2.9@J%#L.6.2.10LRSSB*6.2.11@B%*%*6.2.12@B&#.6.2.13#a@#B:0.5pL.2pL.10pL,20pL,100pL,200pL,lOOOpL.6.3f#SMR%%%29h,E%SzPJN%M*8%SjiYkt%BJ06.3.1%QM&:Ek3$GB/T6682+--&*b9B&.6.3-2RNaseB#3#:H%%Sf:&&3&7fCAji*#37kRNAseAi@rft;T~@,#3Rh3tH%B%10g/~,*%h3;--20.c~a,sse~iiar*ao6.3.3CTAB@r$*:CTAB20g/L,sk@j1.4mol/L,Tris0.1mol/L,Na2EDTA0.02mol/L,Bj2&8413SSkMBTpHH8.0.6.3.4&!#:db$$BBI.AbFI-%HH@ia3%h%$&BIL12DNA.6.3.5%7)cL~.#~iS%i$'~~fp106.3.6TE@@%:Tris0.010mol/L,Na,EDTA1.0mmol/L,H&j@3~~kMRTpH38.0.6.3.7PCRt&Bj&($i$'Mg++PCREG@i,dATP.dTTP.dCTP.dGTP.TagB$.).6.3.831%~sm@i~~~Han~ewfiasm~~~as~~a~mfi~m3I~~~1Jasloa;Iuia4autna~lse~~~~gi~~jiam1~1~~~gi;g~~~1#a~]r~~&NLBPE3-PEPcase1-ectinIVRZElNtRNALeuCaMV35SFMV35SNOSNPTllPATBAR#:SN/T1196,SN/T1197@SN/T1199.41CFFPJiE:5'-ccagttcttggagccgcttga-3'E:5'-aagggccagtccaaatgcaga-3'iE:5'-gccctctactc~acccccatce3'Iji:5'-gcccatctgcaagcctttttgtg-3'iE:5'-cctcctcgggaaagttacaa-3'Iji:5'-gggcatagaagglgaagtl-3'iE:5'-ccgctgtatcacaagggctggtacc-3'Iji:5'-ggagcccgtgtagagcatgacgate3'iE:5'-tgaacccatgcatgcagt-3'Iji:5'-ggcaagaccattggtga-3'E:5'-cgaaatcggtagacgctacg-3'Iji:5'-tt~cattga~tctct~cacct-3'iE:5'-tcatcccttacgtcagtggag-3'Ijl:5'-ccatcattgcgataaaggaaa-3'iE:5'-aagacatccaccgaagactta-3'Iji:5'-aggacagctcttttccacgtt-3'iE:5-aagcctctlacaaggtctlg-3'Iji:5'-ctgctcgatgttgacaag-3'iE:5'-gaatcctgttgccggtcttg-3'Iji:5'-ttatcctagtttgcgcgcta-3'iE:5'-atcgttcaaacatttggca-3'Iji:5'-attgcgggactctaatcata-3'iE:5'-ggatctcctgtcatct-3'Iji:5'-gatcatcctgatcgae3'jE:5'-aggatctcgtcgtgacccat-3'Iji:5'-gcacgaggaagcggtca-3'jE:5'-gtcgacatgtctccggagtlg-3'Iji:5'-gcaaccaaccaagggtatc-3'iE:5'-a~aa~cacggtcaacttcc-3'Iji:5'-actcggccgtccagtcgta-3'L~M%R.~H.~%.RH%J~~#~uI~%~H~IP+R~&~~BHPCRP4?bk/I\/bp121118162226173180165210196180165173183191175&B&Brn%R~~2&ElkaPl&&L%l5%PlBSB&#at%#BYh--9'maa%HaAPa%%aMHrnL2%845k;kHRoundupReady).H8(Bt176.Btll,Mon810,T14/25).%8RH%E#rnrnL~S&B%I%HN~@H%E#maL2sffHARoundupReady).35At(Htll,CA21,).a~H+mH%EmmaL2%2&Brn%H8M*F%@#maL2%3&Ed;kQ.H%.3t8%#3%R#B(1aL2$$sBla%Ha.3*(Bt176)%E#~i1hb53LH&H5l%@flXSYlT1202.6.4@iW$fist6.4.1ef@#P&#QZE2~I~RJ~~+~JIIA2X0.5mL&H*B400pLTES)rp#&,@@l%EE5min9SfTR~t13000r/min3min,&BkEaflB,E;~&I$ll4--3i:+fiIIA2X0.5mL&Ha,?&rL~,&l%kEil4flBE,EX=SZ&.R7kNHTfRDNA.6.4.2fiMf&MEPDNA&a@!6.4.2.1CTABS?E$l#&k@%l400rLb!l%k#l+,fiUA400rLCTAB@%#&%I2rLRNaseB%#&@?&1%1s-3s,fT65TZkB30minrhRA%#%241ZS,Eff~/;WIftBB,$3$!~~BftG$iBi5min,13000r/rnin$PTR~Lh5min(t~%k##E%%&.%Z&4W);%t3#85+kR@?TZ31.5mL?&JL~@;T%~~&J~@$~~uA1rg-2rgB#(Carrier).#fi~A0.6Vol~R~~H;W~iS,~-fP1~~ZT-~OC#$~1h--2hE$2DNA113000r/min$@&&10min,#&k@j#&;H70%ZE#%%BE=&,+B%EQ,@~RJ~W$~UA30rL-50rL0.1XTE!l#)rp#&%@E$2,4C4%@4&H.6.4.2.2iit#fl&S&BST5HB%RBHSB%DNA@@S$J&,@HW#BS$J&H%fi%g+%fi,%DNAE~W,RBUA~HM#RBZ~~T~HM~B+DNAH~~IE~E,%B~BM,JS~~HSRI&%o6.4.3fMf&SFP#il[g&ZfSDNAEf&!&a6.4.3.1g9b33k3kBS%HSkfi5+%%B%NM&HM13+#ArBrHDNA%fiZS,fiNnOD@%Q%E!#&8:Ht$@,k!.a.flk#J-)%%@%.3$M&@#ZEHa#%ag2pg/mId--50pg/mL,OD@(opticaldensity)!$%%0.05-1t?JEFililptl#6DNA%@?Bildd~%H~#,~A%3b3).%%IfEitH~b~+.T260nm4k1~#%~&~,10D260m=50pg/mLB!@DNA338pg/mL@@DNA.PCR9DNA&$#&@OD260-/ODZBO,,,,,&@%1.7-2.0,6.4.3.2IQBsBlrMSaidPCRffM~~~VMU~##~RNHhtJSBo6.4.4PCR#WEE6.4.4