*4.6. Total Cellular DNA Extraction and PCR Analyses*

Total cellular DNA was isolated from leaf tissues of antibiotic-resistant sugarcane clones as well as from nontransformed plants. PCR analysis was carried out to validate transgene integration. *Taq* DNA polymerase (Fermentas, Hanover, MD, USA) and platinum *Taq* polymerase (Fermentas, Hanover, MD, USA) were used with 100–300 ng of genomic DNA as a template. The *gfp* gene was amplified with primers P5 (5- -CCA TGG CTA GTA AAG GAG AA-3- ) and P6 (5- -TTA TTT GTA TAG TTC ATC CA-3- ). The left border fragment was amplified with primers P1 (5- -GAT ATC AAA ACC CGT CCT CAG TTC GGA TTG C-3- ) and P8 (5- -GGG CTG ATA CTG GGC CGG CAG G-3- ).
