RescueMu plasmid library plate screening for a gene with multiple insertion sites. (a) Schematic diagrams of a RescueMu insertion hotspot: demonstration of the assembly of flanking genomic sequences; locations and directions of all primers used in this study; EST alignment to genomic sequence assembly showing introns. (b) An ethidium bromide-stained agarose gel of the PCR products from columns 1 and 22 and row 42 plasmid libraries, using primer pair 2 + L. (c) An ethidium bromide-stained agarose gel of the PCR products with leaf DNA extracted from G42-22(x) progeny 1 to 8, using primer pair 3 + 6. (d) An ethidium bromide-stained agarose gel of the PCR products with the same DNA used in (c), except using primer pair 3 + L (column B is blank). (e) NcoI-digested DNA blot from plants 1 and 3 to 8 probed with a fragment spanning a 0.6-kb PCR product amplified with primer pair 1 + 5. (f) Phenotypes at several developmental stages (from left to right): 10-day-old seedlings (1 to 10 from left to right) of the G42-22(x) progeny; a side-by-side comparison of plants 5 and 6 at 10 days, including their root mass; adult plants at 1 month showing plant 5 in the foreground of the picture with two siblings on either side; a close-up of the plant 5 adult leaf phenotype.