TAIL-PCR for the recovery of Tnt1 flanking sequences in chickpea: a tool for functional genomics studies

Abstract

Thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR) is a powerful technique for amplifying genomic regions flanking Tnt1-retortransposon insertions in plants. Here, we present a TAIL-PCR protocol for amplifying Tnt1-flanking genomic sequences in chickpea using Tnt1-transformed hairy roots as the starting material. The amplified products can be cloned and sequenced for the precise mapping of Tnt1-integration sites in the chickpea genome. This method enables the functional characterization of chickpea genes governing root-specific traits and can be easily adapted for flanking sequence tag recovery in chickpea Tnt1-mutant populations.