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Targeted MinION sequencing of transgenes.

Abstract：

: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.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Boutigny AL,Fioriti F,Rolland M

doi

10.1038/s41598-020-71614-6

subject

Has Abstract

pub_date

2020-09-15 00:00:00

pages

15144

issue

1

issn

2045-2322

pii

10.1038/s41598-020-71614-6

journal_volume

10

pub_type

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