Modification of TAIL-PCR and NBS-RGC5 gene flanking sequence cloning from banana

Fusarium wilt disease is a destructive fungus disease for banana, it has serious negative impact on banana planting and industrial development, so it is significant to research the resistant gene related to banana wilt disease. In this article, part of gene sequence of Nucleotide Binding Site type of Resistant Gene (NBS-RGC5) from Brazil Banana (Musa acuminate ssp. Cavendish AAA)was isolated using modified TAIL-PCR method.Compared with the original protocol,there were two main modifications of the TAIL-PCR：(1)Instead of the single of the degenerate primer, the combination of two degenerate primers was used in the modified TAIL-PCR.(2)The annealing temperature in the 15 super cycles in the Primary PCR was set differently.The final results demonstrated that the modified of TAIL-PCR was an instant and efficient method to clone the flanking sequences from known region.