Genomic DNA Extraction and PCR Analysis

Genomic DNA was extracted from wild-type and putative transformants by using an RBC Real Genomics Genomic DNA Extraction Kit (YGP50, RBCBioscience) and used as a template for PCR analysis to test the presence for GFP using the following specific primers: GFP-F: 5′-CAAGGGCGAGGAGCTGTT-3′; and GFP-R: 5′-CTTGTACAGCTCGTCCATGC-3′. The chloroplast marker atpF-atpH was amplified as an endogenous control [43 (link)]. The PCR program was as follows: 94 °C denaturation for 10 min, followed by 30 cycles at 94 °C for 30 s, 50 °C for 30 s, and 72 °C for 40 s, and a final extension at 72 °C for 7 min. The PCR products were resolved by electrophoresis in 1.2 % (w/v) agarose gel and stained with SafeView™ Classic (Applied Biological Materials Inc., Richmond, BC, Canada).