Genomic sequences flanking the T-DNA insertions (FST: Flanking Sequence Tags) have been determined for all the T-DNA lines in the collection. 46,236 flanking sequence tags (FST) have been identified, they are available in the database Flagdb++, as well as SIGnAL and TAIR.
To confirm a T-DNA insertion, PCR amplify the genomic DNA of the mutant line using one primer from the T-DNA border and one primer specific for the genomic sequence. Since rearrangements occur during the translocation event, it is important that gene specific primers are designed directly from the FST.
If the FST was amplified with the right border (RB) of the T-DNA, you can use the primers
or RB4: 5'-TCACGGGTTGGGGTTTCTACAGGAC-3'
If the FST was amplified with the left border (LB) of the T-DNA, you can use the primers
or LB4: 5'-CGTGTGCCAGGTGCCCACGGAATAGT-3'
As our T-DNA insertion lines are in the Ws (Wassilewskija) background, use Ws, not Col-0, as a control for gene amplification and phenotyping.
Contact us if you have any questions. Please always include your client code, and include your order number if your inquiry concerns an order.