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O.5mmol/Lofeachouter F3andB3.3.0mmol/Lofeachinner FIFand
primers primers
DNA 0mM 0
BIP,2.5pL凰f polymerasebuffer(20mM%s-HCl(pH8.8,250C),1KCl,1
mM X-l mmol/L mmol/Lof
mM(NI-h)2S04,2MgS04,0.1%TritonOO),0.5 Mg十,1.5
each of8UBstDNA 50 the
deoxynucleosidetriphosphate,0.7pL polymerase,andng
DNA.Themixturewasincubatedat650Cfor60minina blockandthen
target heating
heatedat800Cfor2mintoterminatethereaction.TheIA^但fordetectionoftheGM
cottonwascarriedoutina oftotalreactionissameasrice 1.0mmol/L
25“L except Mg什,
and5mrnol/L were ina size
dNTPs.LAN口products 2%agarose
electrophoresed gel.The
ofthe ofthe
fragmentLAMP isin withthe ladderlike
productgoodagreementpredicted
size.
ThedetectionlimitsofPCRandLAMP theDNA extracted
assaysusing templates
withCTABmethodwere0.1%and0.0 IAMP wasfoundtobe
1%.respectively.Theassay
1O.foldmoresensitivethanthe仃aditionalPCR addition.thereweredifferent
assay.In
threekindsofnon-GM kindsof ofnon-GM
samples(contain rice,three cotton,twokinds
kindsofnon—GM 3kindsof Btstrains
soybcan,two tomato,and positive containingcrylA
bedetected
PCRandI.AMP inordertoevaluatethe of
gene)to by respectively
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