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. 2005 Oct;17(10):2619–2632. doi: 10.1105/tpc.105.033506

Figure 7.

Figure 7.

Variation in G. max F3H Genomic Amplification between Lines Varying at the Wp Locus.

Ethidium bromide–stained gels showing the results of genomic PCR amplification using the following.

(A) The 38F and 1357R primer set. A 3.5-kb fragment was amplified in two lines with the Wp allele (LN89-5320-6 [Wp] and RM30 [Wp]) (Table 1). By contrast, no significant amplification occurred in the mutant isolines LN89-5320-8-53 (wpm) and LN89-5322-2 (wp).

(B) The 7F and 1428R primer set. A 3.5-kb fragment was amplified in the LN89-5320-6 (Wp) line, and an additional 2.7-kb fragment was amplified in both the LN89-5320-6 (Wp) and the mutant isoline, LN89-5322-2 (wp).

(C) The 7F and 1428R primer set and higher annealing temperature (58°C). This change favored the amplification of the 3.5-kb genomic fragment in the LN89-5320-6 (Wp) line.

(D) The 38F and 1428R primers and new set of PCR conditions that favor amplification of larger fragments (see Methods). A 3.5-kb fragment was amplified in the LN89-5320-6 (Wp) line, while a 9.2-kb fragment was amplified in the mutant LN89-5322-2 (wp) isoline.