Figure 1.

Main steps of cassette-PCR genome walking (GW) and pyrosequencing association strategies. The original strategy reported by Wang et al. [6] (A) and its improvement by the adoption of barcoded primers [13] (B) are illustrated. Blue and white bars correspond to known and unknown sequences, respectively. A black horizontal arrow indicates adapter specific primers; a blue arrow represents primers designed in the known region. The symbol indicates a restriction site. Barcodes and 454 sequencing traits of longer amplification primers are indicated in the figure.