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Figure 2 | BMC Plant Biology

Figure 2

From: Discovery of chemically induced mutations in rice by TILLING

Figure 2

Mutation discovery in eightfold pools of mutagenized individuals arrayed in a 2-D scheme. Mutation discovery in eightfold pools of mutagenized individuals arrayed in a two-dimensional format. Top: A two dimensional pooling scheme is used whereby sixty-four samples from individual rice plants are first arrayed in an eight by eight grid. Samples from a common column are pooled in the first dimension and from a common row in the second dimension. Row pools and column pools are arrayed on adjacent lanes of an assay plate. Bottom: IRDye 700 (left) and IRDye 800 (right) gel images from the same sixteen lanes of a ninety-six lane TILLING assay for mutations in the 1,167 base pair OsDREB rice target. True mutations (boxed) produce a band in the column and row pool of each IRDye image, allowing the unique individual harboring the mutation to be determined in a single gel run (marked in yellow on the top diagram). Bands resulting from the same mutation are boxed a similar color. The molecular weight of the mutant band in the IRDye 700 image plus the molecular weight of the band in the IRDye 800 image adds up to the molecular weight of the full length PCR product. To the left of the images a virtual molecular weight ladder generated by the GelBuddy gel analysis program is shown. Next to this is the gene model (lines represent introns, boxes represent exons) that was generated when target primers were chosen using the CODDLe program.

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