Identification of T-DNA structure and insertion site in transgenic crops using targeted capture sequencing

The commercialization of GE crops requires a rigorous safety assessment, which includes precise DNA level characterization inserted T-DNA. In the past, several strategies have been developed for identifying T-DNA insertion sites including, Southern blot and different PCR-based methods. However, these methods are often challenging to scale up screening dozens transgenic events with complex genomes, like potato. Here, we report using target capture sequencing (TCS) characterize structure 34 in This is an 18 kb fragment between left right borders carries three resistance (R) genes ( RB , Rpi-blb2 Rpi-vnt1.1 genes) that result complete late blight disease. Using TCS, obtained high sequence read coverage within junction regions. We identified breakpoints on either ends 85% events. About 74% had their 3 R gene sequences intact. flanking were from potato genome half events, about third (11) single mapped into genome, five do not interrupt existing gene. TCS results confirmed PCR Sanger 6 best representing 20% suitable regulatory approval. These demonstrate wide applicability crops.