Chromosome 7: TG572, 48cM

TG572 RFLP Probe: Five primers were designed around the RFLP probe on chromosome 7: P572F1, P572F2, P572F3, P572R1, and P572R2 (Table 1). All possible primer combinations produced a strong band with Heinz 1706 DNA between 600 bp and 1000 bp (Fig. 2). All of the primer combinations produced single bands with the exception of P572F2/P572R1, which produced two bands one at 400 bp and one at 900 bp. The P572F1/P572R2 primer combination produced a strong single band at 800bp and was chosen for use with additional genotypes. Some primer combinations produced larger and stronger single bands, but PCR fragments larger than 900 bp are often difficult to sequence with only one primer pair and thus these primer pairs were passed over. Fig. 2: Agarose gel of the PCR reactions with the TG572 primers and Heinz 1706 DNA. lane 1, 100-bp ladder; lane 2, P572F1/P572R1; lane 3, P572F1/P572R2; lane 4, P572F2/P572R1, lane 5, P572F2/P572R2; lane 6, P572F3/P572R1; lane 7, P572F3/P572R2, lane 8, positive control P301F3/P301R2; lane 9, water. The P572F1/P572R2 primer pair was chosen for sequencing reactions due to its size and the brightness of its band. Arrow marks the 700bp fragment.