SNT 1202-2003 食品中转基因植物成分定性PCR检测方法

#%S*S&NB%B*ZtBPCR&jJ!g:gProtocolofthequalitativepolymerasechainreactionfordetectinggeneticallymodifiedplantcomponentsinfood~3n~~f+t:~)1m~kaa*k@m~1milfiljt%*h%m%&~nmfl:.asrsssmxFa+le.mmi%H#&@t~&#~R&~@)~#t5%~4BH;f.;4%~t%TmtBfiRB2P;Z~B&B~~%3iWf%ABRf@!HBBAt+;9~%E*eRR+%f:~tgl5lMFf]R~1~~~zb:~H~*~~.GB/T6682j)$i%&SH7k%#j#IBtk&(eqvIS03696:1987)SNIT1193J#B@!&%&3Z&*BjkSNIT1194M@B#~#I%&~&~@B!!~#~#~!#-)~%SN/T1195hEi+$$SL!4&3)Z?4I'CRUB!!YJ'&SN/T119639%PCR%%!!h?&SN/T1197?h%RR&B3&3)2#PCR3$ildhk&SN/T1199#l%+R&B3&3)PCR&@I%-&SN/T1204t$~B#%fi~SC~&~%3&~r3t3)Td'U~%XPCR%&%t%?f?&TPl@%%&HT*&%*3.1PCRIpolymerasechainreaction,m%PCR.3.2DNA1de~x~ribonucleicacid,%%@@j&@.3.3dNTP:deoxyribonucleosidetriphosphate,8#$t%3?@@.3.4dATPldeoxyadenosinetr~phosphate~Llk!%IlTT@@.3.5dCTP:deoxycytidinetriph~sphate,a#SmBE-R@~3.6dGTPIdeoxyguanosinetriphosphate,R#$€~~7@@3.7dUTPdeox~ur~dinetriphosphate,Jl$!$G$e3.aft@3.8bpbasepair,@SN.3.9TagBrDNAJ@*m.3.10'I'rls,tris(hydroxymethyl)amlnoniethanc,!-:(%!pg)&,%V%$.,3.11TE:T~I~-CI.EDTA$~!~~~$..3.12I.ectinl&44&l%~.3.13ZEINi#&%B$#E$.3.14~~3-~~~case1~hos~hoenil~~ruvate-carboxylahc,b#@~~~aj~@;)fat.(t:~.'3.15~atat~n,~##&ZE%Z?lAPalatinBH#&jttlR%q%BHli4%%fl:e3.16tRNA~eu~&kQdWflW#4f!il?il.3.1718srRNAl&&%@118sRNA%a.3.18CaMV35S135Spromoterfromcauliflowermosaicvirus,?f€aT8@%%lJt$i%l8J35SBaT.3.19NOS,terminatorofnopalinesynthasegene,MRB~-&lilt~~~~k~.3.20FMV35S135Spromoterfromamodifiedfigwortmosaicvirus(caulimovirusgroup),g8ZR-tR*35saa~.3.21NPTII:neomycin-3'-phoephotrmsforasegene,#$#-3'-@@R@BS88.3.22BAR:phosphinothricinacetyltransferasegenefromBacillusamyloliquefacions,?#@TfimBacillusamyloliquefacisnsVTj#(Ptl%%@E#SBj.3,23PATIphosphinothricinacetyltransferasegene,9TM2E?Rl$~&B.3.24COXIglyphosateoxidoreductaaegene,W€FM~gtEHll#ftlE~.3.25CryIA(b)lasyntheticgeneencodesthefiret648aminoacids,insecticidal-activeftruncatedproductidenticaltothatofcryIA(b)geneofBacillusthuringiensissubsp,%2$?$84HB%&isI3crylA(b)SB.3.26Cry3AlB,thuringiensissubsp,Tenebrionis(B,t,t,)strainQI256-82B&@sfi,&##k@*#bjQ&Il~*aEfl&(Coloradopotatobeetlelarvae).3.27EpSPSI5-enolpyruvylehikimate-3-phosphatesynthaeegene,5-%9?@-3-@@.ftj%jtSB9..3.28PVY-cplpotatoYviruscoatprotein,$@aY#8%4/9!iSb.3.29PGIp~l~galacturonase,%#$SL#km@$I$f&&qRSB&3);BB@a8mmmffl!#BBiBR8SNIT1193l8J%4Se6.1Q(fPWdiBS&@iDNAg,HNrf&WM@Be$ABCI4bWS$IM#B#flR#3lC,NUPCR~$,DNAHWB~PCRr)11~~~~,~~[lfia~fi~~~e1$egae:~~itt3f..6.2PZNlll6.2.1PCR%@@fi.6.2.2DNANBfi.6.2.3B?9lt#b.6.2.4E$%!X.6.2.58W9i%@%&o6.2.6#iRB$IR.6.207MB!BB3)fi.oY*6.2.0&i$*l&s~ba,6S/j\B$f~b#l,Mini/.6.2.9@B%$#,@@@W;%#B.6.2.104$7Jc#,~Xa[#k#~6.2.11#11]bfon.6,'2.12&ASB#.26.2.13s?J#~,~B;56B~M%~~H#~@&~@f#~6.2.14$FRY.6.2.15BB&#!qPC~@@%fB%l-6.2.16#*@%#80.5pL.2pL.10pL.20pL.100pL.200pL.1000PI..6.324iitIf'dRBGMBM~Eff4~~1~%+BTM&!k21:&fpla6.3.1%%R&~&flf:GB/T6682q-%7JCb9B#fO6.3.2%llBMF.6.3.3RNaseH%%tli20@WmgD~TICI~HMSJYI%&-&FF&B!%~~~~#IRNAsc-AMl@B%10g/I~~+%fiT-2O~#@~%%EB%H~6.3.4CTAB%@@:CTA1320g/I.,#&+&1.4n~ol/I-~l'ris0.1rnol/L,NalEDTAO.02mol/I.,Rj&@&%$f,ikQltjNPipH358.0.6.3.5CTAHW,s#iICTAB5g/L,%kM0.04mol/I..6.3.6#,4&@9@@!ia-#k@jl.,2mol/l.,6.3.7BifiiYlKKRIIQMWlf20mg/mi..%T1flS~kl~~&bN%lMWT-20'CC',B%lZ$Z%fil4.6.3.0B$#t~~~&4F4.4~~-##'b9&~~3#~~~~DNAe6.3.9TEBNj&tTris0.010rnol/l.,Na,EDTAO.001mol/LlJa&~&S,lTB-#k~i141~pHH8.0.6.3.10%7)CtS.WTJ4W%#i.Uliieffi1.6.3.11&!iEl&DNA@RlrC%!f.6.3.12PCRa3PJ&lrqMgfPCR~#@.dATP.dTTP,dCTP.dGTP,TagA.6.3.13Q#br#@N%(I,oadingHuffer).6.3.1451%e~~s~~~~+~a~~mama~*~ba~wfi~~m~~~~i:~das1v~xx~m~w~ffiNbUIb9.%tpa+#ttk%+IBINSB@Wr3I??aFFM!&SN/T1196-2003$USN/T1197--2003.1&NJ*rffr$PlB$5BitW1@*4E@SNH3I3$#Pl48tiU!lM26!3iCaMV35S1FMV35Sa.NOS'NPTII'PATm35S/PetuCaMV35YCTPolWSPSCOXCP4-EPSPS($#IBARLBB#R9bm$h%NmaHd9S$19i-HW#J%mfinWHlilfiBk1~~%~BEB!Eib!lJ&Bis8fi&WbQbB;8H%F!B1?1H%BBkEL(RoundupReady),%~(Bt176,Btll,Mon8lO,T14/25).%IH~,?#%Zenecaa).$@*%retNm*6aU@S!A%R%l,$##(Ne-wLeafY.Newl.esfPlus);#di)fa%BHBrJlRaH+$&EkIi(RoundupReady).sX(Btll,GA21r).%#%I.#%(Zeneca0,4+$#%JX#~-BracZlR&faB5&\4+4f*#RJIR;jl~PIBP*WI~z~R%~~~A~~x,AIR.I~B%IB~W~i?~-btrsAH%%Rkl2%Et#bQf&cZlR~BB%~~%~~~l?t&URi%B#EHMttll3i~XBl176,CBH-35l)%@#+M.BtWIYl@#3U:5'-tcatcccttacgtcagtggag-3'Iji15'-ccatcattgcgataaaggaea-3'E,5'-aagacatccaccgaagactte-3'bi15'-aggacagctcttttccacgtt-3'iEI5'-nagcctcaacaaggtcsg-3'gI5'-ctgctcgatgttgacaag-3'E15'-gaatcctgttgccggtcttg-3'15'-ttatcctagtttgcgcgct&3'E5'-atcgttcaaacatttggca-3'R15'-attgcgggactctaatcata-3'EI5'-ggatctcctgtcatct-3'g15'-gatcatcctgatcgac3'--iE5'-sggatctcgtcgtgscccat-3*,5'-gcacgnggaagcggtca-3'iEI5'-gtcgacatgtctccggagag-3'15J-gcaaccaaccaagggtatc-3'e15'-tgatgtsatatctccactgacg-3'15'-tgtatcccttgagccstgttgF3'iE15'-ccactatccttcgcaagacccttcc-3'15'-ttgtatcccttgegccatgttgt-3'5'-gtcttcgtgttgctggaaccgtt-3'fi15'-gaactggcaggagcgagagct-3'IE,5'-gacttgcgtgttcgttcttc-3'&15'-aacaccgttgagcttgegac-3'H5'-acasgcacggtceacttcr3'5'-ectcggccgtccagtcgta-3'PCRPQk/l\/bp165210196180165173183191171120120204175I5'-ggatccttagaagcatctagt-3'PC/NOS35062:5'-catcgcaagaccggcnacag-3'RB1114581.#&@f%~ilmilr#ttl383bp%I180bpik4+;l;~;k/~\I)NA~+;.ee:.~c%ss8.n~n~mrq3a3I,PtttlDNA6.4&M?b%6.4.1#Y#H#J#&H%tfill&&&%#3b!J%R