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Information Resource
Record information and status
Record ID
115854
Status
Published
Date of creation
2020-12-17 15:09 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-12-17 15:09 UTC (austein.mcloughlin@cbd.int)

General Information
Title
Targeted MinION sequencing of transgenes
Author
Anne-Laure Boutigny, Florent Fioriti, Mathieu Rolland
Language(s)
  • English
Publication date
2020-09-15
Subject
Summary, abstract or table of contents
The presence of genetically modified organisms (GMO) is commonly assessed using real-time PCR methods targeting the most common transgenic elements found in GMOs. Once the presence of GM material has been established using these screening methods, GMOs are further identified using a battery of real-time PCR methods, each being specific of one GM event and usually targeting the junction of the plant genome and of the transgenic DNA insert. If, using these specific methods, no GMO could be identified, the presence of an unauthorized GMO is suspected. In this context, the aim of this work was to develop a fast and simple method to obtain the sequence of the transgene and of its junction with plant DNA, with the presence of a screening sequence as only prior knowledge. An unauthorized GM petunia, recently found on the French market, was used as template during the development of this new molecular tool. The innovative proposed protocol is based on the circularization of fragmented DNA followed by the amplification of the transgene and of its flanking regions using long-range inverse PCR. Sequencing was performed using the Oxford Nanopore MinION technology and a bioinformatic pipeline was developed.
Thematic areas
Information on Organisms or LMOs
Gene(s) identification
CaMV 35S promoter - Cauliflower mosaic virus - CaMV
Dihydroflavonol-4-reductase - Zea mays - Maize, Corn, MAIZE
Changes in quality and/or metabolite content - Pigmentation / Coloration
CaMV 35S terminator - Cauliflower mosaic virus - CaMV
Nopaline Synthase Gene Promoter - Agrobacterium tumefaciens - Agrobacterium
Neomycin Phosphotransferase II - Escherichia coli - ECOLX
Resistance to antibiotics - Kanamycin
Nopaline Synthase Gene Terminator - Agrobacterium tumefaciens - Agrobacterium
Additional Information
Type of resource
  • Article (journal / magazine / newspaper)
Identifier
https://doi.org/10.1038/s41598-020-71614-6
Publisher and its location
Scientific Reports
Springer Nature
Rights
Open-access
Format
PDF - 1.02 MB (7 pages)
Source
Article in online journal
Keywords and any other relevant information
Sequencing; bioinformatics; inverse PCR; unauthorized; transgenes