Efficient Amplification of Insert End Sequences from Bacterial Artificial Chromosome Clones by Thermal Asymmetric Interlaced PCR

TAIL-PCR is a powerful tool for the recovery of DNA fragments adjacent to known sequences. A protocol is presented for the amplification of insert end sequences from bacterial artificial chromosome clones using TAIL-PCR. The amplified products are suitable as probes for chromosome walking and genome mapping and as templates for direct sequencing. The protocol has been used in rice genome studies. Abbreviations: AD primer, arbitrary degenerate primer; BAC, bacterial artificial chromosome.