FIGURE 6.

Identification of transgene integration by genome walk technique. Steps involved in genome walk technique. Restriction digestion of genomic DNA of ‘D3’ transgenic line (high glyphosate tolerant) with BglII enzyme (A). Initial amplification of transgene integrated product using adaptor specific and T-DNA specific primers (B). PCR amplification of transgene integrated product using first PCR product as template (C). Confirmation of genome integrated T-DNA sequence cloned in TOPO vector by colony PCR (D). Identification of transgene integration site in the rice genome using BLAST program (E).