Figure 4

Eco-TILLING using ENDO1. (a) Schematic representation of a 456 bp genomic DNA fragment of Arabidopsis thaliana showing two sequence polymorphisms, 3-base INDEL and the G to A substitution, between Columbia and Landsberg ecospecies.(b) The 456 DNA fragment was PCR amplified using the primers indicated in Table 1. Heteroduplex DNA is prepared by mixing equimolar amounts of the two PCR products, generated from Columbia and Landsberg ecospecies, heating to denature the DNA followed by cooling to renature it. Lane 1 indicates heteroduplex DNA. Lane 2 and 3 indicate homoduplex DNA generated from Columbia and Landsberg ecospecies, respectively. Homo and heteroduplex DNAs were digested with ENDO1 and analyzed on LI-COR4300 DNA analyzer. Expected band sizes are indicated on the left of the panel. (c) DNA sequencing chromatograms surrounding the 3-base INDEL (Left panel and underlined) and the G to A substitution (Right panel and underlined) between Columbia (upper sequence) and Landsberg (lower sequence) ecospecies.