《SNT1195-2003-大豆中转基因成分的定性PCR检测方法》.pdfVIP

Protocol of the qualitative polymerase chain reaction [PCR) for detecting genetically modified component in soybeans T P J * i a . B X ~ ~ r $ i i ) ~ R 3 = * ~ ~ 0 3. 1 %a aft genetically modified component ~ C ~ * A + a ~ - m ~ s m ~ s ~ ~ ~ m a L % m ~ p r l . 3.2 NefiaX6i!iploymerase chaln mctlon (PCR) ~ ~ ~ F F ~ ~ 9 % %DNA~~ B ~~ #~ R# ~~%B ,#%#fE.$iJT9WtM~ E~ ~ ~ T ifMM*4I@%Si;U%.$i DNA ~%iZFI#.fl~-X+E~PJ,]~ATlii~X@~,##DNA BM6HTUDH~RR#BbSfl(dNTP)%@,@3l~~UZ~$tlr#ZRELSZ%%~~kN%f$ -ms,@ar#@sB*ufiN%Br#o 3 . 3 @*#) 3 . 3 . 1 Lectin~4$#B3tSA ,%kH*9j)%ZftN CIJlSB. 3.3.2 CaMV 35Si35S promoter from cauliflower mosaic virus,#EtEiBS#t$E4~3 5 S B 3 3 . 3. 3 . 3 NOSrTerminator of nopaline synthase gene from Agrobarterium tumefaciens ,sSTffg MM117i#i-B4iB@It:3D 3 . 3 . 4 CP4 EPSPS: 5-enolpyruvylshikimate3-phosphate synthase gene, 5-#t$WBW @%gZt4t-3-@@ QdBFiB;I* 3 . 3 . 5 RRS:roundup ready soybean,3iH Monsanto B k B , R A f i S 3 f i B % CaMV35Sh33113,NOSL3@ CP4-EPSPS2iW%B.. 1 6.1 81 !@ RRS %MA%BSB%+%RH9JBSBBPUIJ.lft3Lt-a@W5I%,tJR PCR ?f%XifS B9E%S$1H@9~fiB%liFB,!%%Q%R%$JE~#an~BI+EB%ffRRS Br3%S89 ern 6 . 2 ii.t%J#lM# B%G29E9fi@MmSBJ%%EB%kS3Ua 6.2. 1 CTAB ,CTAB 20g/L, Tris-HCI 0.1 mol/L(pH8. 0) ,EDTA 0.02 mol/L. 6. 2. 2 Tris fQ$UiaBB. 6 . 2 . 3 f a R F ~ : R f i B q r ( 2 4 1). 6.2.4 SWB. 6 . 2 . 5 70%ZfJ$. 6.2.6 T E @ @ ( l o mmol/L Tris.1 mmol/I, EDTA,pH8.0). 6.2.7 RNA m%% :5pg/pI.. 6 . 2 . 8 IOXPCR @G@. 6.2.9 aB(MgC12) :25 mrnol/L. 6.2.10 dNTP(dATP,dCTP,dGTP, d U T P ) B a I 2 . 5 mrnol/L. 6.2.11 Taql$$:5U/pL. 6.2.12 51:%45Bk~tS~~B~9EBS@slU1~fBHfli;l8B3~l%S1. sW 1 SSHkHMtlASm*U9EISHM91%A:W U 1 I B