Assessment of the genetic stability of GMOs with a detailed examination of MON810 using Scorpion probes

Neumann, Georg; Brandes, Christian; Joachimsthaler, Alexandra; Hochegger, Rupert

doi:10.1007/s00217-011-1487-8

Cited by 9 publications

(16 citation statements)

References 42 publications

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“…In contrast to the earlier investigations on MON810 [ 20 , 32 ], our study examined MON810 in a stacked event. Combinations of transgenic traits increase the complexity of risk assessment because more undesired scenarios may occur.…”

Section: Resultsmentioning

confidence: 99%

“…These tests can be performed readily and are cost effective. The characterization of the Scorpion primers used for the analysis of GMOs has been described by Neumann et al [ 32 ] and Thelwell et al [ 39 ]. Madi et al [ 31 ] have shown that Scorpion PCR combined with HRM analysis is a highly sensitive method for the detection of DNA alterations, including SNPs.…”

Section: Resultsmentioning

confidence: 99%

“…For the investigation of 5' junction of MON810 (5'-MON810), a stacked GM maize variety (MON88017 × MON810) containing three genetic elements was used. The analyzed target region has been described by Neumann et al [ 32 ] and the primers are shown in Table 1 . It was possible to test this region using Scorpion PCR [ 32 ] and HRM analysis.…”

Section: Resultsmentioning

confidence: 99%

“…Genomic samples can be sequenced directly or GMOs can be analyzed by methods such as Sensitive Capillary Electrophoresis (CSCE) or long-range polymerase chain reaction (PCR). Further, real-time PCR employing Scorpion primers for amplification or high resolution melting (HRM) analysis can be used to distinguish between different alleles or to filter out mutated samples [ 31 , 32 ].…”

Section: Introductionmentioning

confidence: 99%

“…Earlier studies have adapted these methods to analyze the genetic stability and organization of transgenic insertion events. In Neumann et al [ 32 ] it could be shown that single nucleotide polymorphism (SNPs) in plasmids can be demonstrated by Scorpion analysis. As a further work for optimizing the method, different alleles of the alcohol dehydrogenase 1 ( ADH1 ) gene in maize were investigated by Madi et al [ 31 ].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis

Ali

Madi

Hochegger

et al. 2014

IJMS

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Genetic mutations must be avoided during the production and use of seeds. In the European Union (EU), Directive 2001/18/EC requires any DNA construct introduced via transformation to be stable. Establishing genetic stability is critical for the approval of genetically modified organisms (GMOs). In this study, genetic stability of two GMOs was examined using high resolution melting (HRM) analysis and real-time polymerase chain reaction (PCR) employing Scorpion primers for amplification. The genetic variability of the transgenic insert and that of the flanking regions in a single oilseed rape variety (GT73) and a stacked maize (MON88017 × MON810) was studied. The GT73 and the 5' region of MON810 showed no instabilities in the examined regions. However; two out of 100 analyzed samples carried a heterozygous point mutation in the 3' region of MON810 in the stacked variety. These results were verified by direct sequencing of the amplified PCR products as well as by sequencing of cloned PCR fragments. The occurrence of the mutation suggests that the 5' region is more suitable than the 3' region for the quantification of MON810. The identification of the single nucleotide polymorphism (SNP) in a stacked event is in contrast to the results of earlier studies of the same MON810 region in a single event where no DNA polymorphism was found.

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Section: Resultsmentioning

confidence: 99%