Figure 3

Hps gene copy number determination by reconstruction hybridization. Soybean (Glycine max cv. Harosoy 63) genomic DNA, or an 11060 bp plasmid carrying a soybean chromosome fragment encoding a single copy of Hps (GenBank accession DQ208939) were digested with Bgl II and separated by agarose gel electrophoresis. The DNA was transferred to Nylon blotting membrane and probed with ³²P labelled Hps cDNA (GenBank accession number AF100159). Shown above each lane is the amount of DNA loaded for the soybean genomic DNA (μg) or for the plasmid standard (ng). The size of the main hybridizing fragment is indicated on the left. Analysis of the band intensities using a phosphoimager results in a calculated value of 27 ± 5 copies of Hps per haploid genome, for the 2.4 kb Bgl II fragment.