BioSample

T-DNA insertions are identified by TDNA-Seq, a method for next-generation sequencing in which pools of T-DNA lines can be sequenced in parallel and then de-convoluted based on their pooling pattern to assign T-DNA insertions back to their originating seed line. To set up the pools, a large numbers of individual TDNA insertion seed lines (10,000) are given a unique four digit number (0000-9999). Based on this identifier a line is placed into one of ten pools (1-10) in each of four superpool sets (plate, row, column and number). All 40 pools (4 superpools x 10 pools) are separately sequenced using a method to enrich for T-DNA/genomic junctions. A T-DNA identified in all four superpools is the unique identifier of the seed line which contained that insert.