Thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR) is an efficient tool for the recovery of DNA fragments adjacent to known sequences. A protocol is presented for the amplification of genomic sequences flanking DNA (T-DNA or transposon) insertions using the TAIL-PCR method. The amplified products are suitable as templates for direct sequencing, or for cloning in vectors. Examples are given in use of the protocol for T-DNA tagging in rice using the pCAMBIA binary vectors.