Patent ID: 8716550

Claim:
Method for the introduction and selection of a specific heritable mutation in M1 plants, comprising: (a) providing a plurality of mutagenized seeds; (b) sowing the mutagenized seeds; (c) growing plants from the seeds; (d) sampling DNA-containing material from a first section of the plant; (e) pooling the DNA-containing material from each first section, wherein said pooled DNA from the combined pools of the first section comprises the first dimension of a N-dimensional grid; (f) isolating DNA from the pool; (g) repeating steps (d)-(f) for a second and any further dimension of the N-dimensional grid for a different section of the plant, wherein each section comprises a different growth and/or developmental stage of the plant and each section is pooled into a plurality of pools and the combination of pools for each section comprises its own dimension in the N-dimensional grid; wherein the combination of pools for the N th section constitutes the N th dimension in the N-dimensional grid; (h) amplifying each pooled DNA with a set of primers that amplify region of the DNA containing the position of the mutation of interest, whereby at least one of the primers contains a pool-specific tag, to obtain reaction products for each of the pools; (i) pooling the reaction products; (j) determining the nucleotide sequence of the reaction products; (k) deconvoluting the sequences from each pool of step (h) using the pool-specific tags to identify each plant in the grid; and (l) selecting an M1 plant having the desired mutation in at least N−1 of the N sampled sections.