The invention relates to a method for producing a chromosome substitution library, comprising: a) producing a population of DH lines from an at least partially heterozygous starting organism; b) identifying a first DH line in the population that has at least one non-recombined chromosome pair; c) identifying a matching second DH line in the population that has at least one non-recombined chromosome pair that is the same as the non-recombined chromosome pair of the first DH line, and at least one non-recombined chromosome pair that is different from the non-recombined chromosome pair of the first DH line; d) selecting the first DH line and the second DH line as a pair of parents, the progeny of which has at least three non-recombined chromosomes, at least two of which form a pair; e) repeating steps b), c) and d) one or more times on the DH population of the heterozygous starting organism, or repeating steps a)-d) on a DH population produced from a progeny plant of a pair of parents selected in step d), until preferably all chromosomes from the selected DH lines are non-recombined and preferably all possible combinations of the parental chromosomes are selected, to assemble a library of chromosome substitution lines. The invention further relates to the libraries thus obtained.