Fig. 2

Molecular markers for vrn-A1, VRN-B1, vrn-D3, and PPD-D1. a PCR marker for vrn-A1. PCR products were amplified using primers VRN-A1F7B/VRN-A1R7, digested with restriction enzyme Sph I, and electrophoresed in a 2% agarose gel. Due to a SNP in exon 7, the vrn-A1a allele showed a band of 197 bp (and a band of 24 bp out of the gel), and the vrn-A1b allele showed a band of 221 bp. b PCR marker for VRN-B1. PCR reactions were performed using the forward primer Intr1/B/F to pair with two reverse primers Intr1/B/R3 and Intr1/B/R4, and PCR products were electrophoresed in a 1% agarose gel. c PCR marker for vrn-D3. PCR products were amplified using primers VRN-D3F6 and VRN-D3R8, digested with restriction enzyme Nco I, and electrophoresed in a 2% agarose gel. d PCR marker for PPD-D1. PCR reactions were performed using the forward primer PPD-D1_F to pair with two reverse primers PPD-D1_R1 and PPD-D1_R2, and PCR products were electrophoresed in a 1% agarose gel. e RT-PCR for vrn-A1 copy number. Genomic DNA samples for Apogee and two plants having the Apogee allele (A) and Overland and two plants the Overland allele (B) were tested. Copy number is shown using the values calculated by the 2^(−ΔΔCT) method, where CT is the threshold cycle. Bar indicates standard error