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Review of peer-reviewed publications Area of the environmental risk assessment: Food/Feed Safety- Molecular Characterization Studies Publication Summary of research and results Conclusion Protection Goal Adverse effects

By Neumann Et Al


Objective: To investigate the genetic stability of MON 810 maize by PCR measurements using Scorpion primers. Experimental Design: 567 individual seeds of MON 810 maize from the same variety were collected from several fields in Germany. DNA alterations amongst individual seeds of MON 810 maize were investigated using Scorpion-PCR by a two-step procedure. First, a large number of individual plant seeds were screened with real-time PCR using event-specific Scorpion primers. Second, samples whose realtime PCR Ct values differed strongly from the mean of the Ct values were screened by SYBR green and sequenced. Briefly, genomic DNA was extracted from seeds and cloned using deferent Scorpion primers for the 5’and the 3 ’ region of the insert. Real-time PCR was conducted under reaction conditions optimized for each Scorpion primer followed by sequencing. Further, reliability of the method was studied by determining several parameters that are required for a validation, such as specificity, cross-reactivity, relative repeatability standard deviation (RSDr) and relative reproducibility standard deviation (RSDR). The results indicate that the MON 810 construct was stable in maize seeds collected from several field trials. Further, several validation parameters studied showed that the Scorpion probe PCR technique is reliable and reproducible

Year: 2013
OAI identifier: oai:CiteSeerX.psu:
Provided by: CiteSeerX
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