Three conditions should ideally be met for a successful mapping cross of three linked genes:
I. The genotype of the organism producing the crossover gametes must be heterozygous at all loci under consideration.
II. The cross must be constructed such that the genotypes of all gametes can be determined directly from observation of the offspring phenotypes.
III. Enough offspring should be produced in the mapping experiment to recover a representative sample of all crossover classes.
Gametes derived from the heterozygous female parent can be determined by examining the phenotypes of the progeny because gametes from the male parent contain all recessive alleles. When a crossover occurs in the female parent, it creates reciprocal crossover classes (both crossover chromosomes appear in the progeny).
In a three-point mapping cross, generally the events corresponding to noncrossover are most numerous and the double crossover events are least numerous. This allows these categories to be identified from the data.