Real-time detection of genetically modified soya using Lightcycler and ABI 7700 platforms with TaqMan, Scorpion, and SYBR Green I chemistries.
A comparative cross platform evaluation of real-time polymerase chain reaction detection of DNA sequences present in Roundup Ready soya was undertaken using the ABI 7700 and Roche Lightcycler detection systems in combination with 3 different detection chemistries: TaqMan, Scorpion primers, and SYBR Green I fluorescent dye. Various copy numbers of a plasmid containing the soya lectin sequence were used to determine the sensitivity and reproducibility of the different technology combinations and to examine both inter and intra machine variability. To examine the relative accuracy of each technology, the genetically modified soya content of baked products containing known amounts of Roundup Ready soya was determined by detection of lectin and the EPSPS transgene. It was determined that the combination of TaqMan detection chemistry and the ABI 7700 platform represented the best method for quantitative detection of genetically modified organisms in terms of both precision and accuracy.
Terry
Catherine F
CF
Bioanalytical Innovations (F008), LGC (Teddington) Ltd., Middlesex, UK. cft@lgc.co.uk
Shanahan
Della J
DJ
Ballam
Lydia D
LD
Harris
Neil
N
McDowell
David G
DG
Parkes
Helen C
HC
D000704:Analysis of Variance; D001483:Base Sequence; D052160:Benzothiazoles; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000032:analysis / Q000235:genetics; D003959:Diamines; D005456:Fluorescent Dyes; D030841:Food, Genetically Modified; D009930:Organic Chemicals; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011804:Quinolines; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000235:genetics*
D052160:Benzothiazoles; D017931:DNA Primers; D018744:DNA, Plant; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I