Positional cloning is a technique that identifies a trait-associated gene based on its location in the genome and involves methods such as linkage analysis, association mapping, and bioinformatics. This approach can be used for gene identification even when little is known about the molecular basis of the trait. Vast majority of traits are regulated by multiple genomic loci called quantitative trait loci (QTL). We describe experimental populations and designs that can be used for positional cloning, including backcrosses, intercrosses, and heterogeneous stocks, and advantages and disadvantages of different approaches. Once the phenotype and genotype of each individual in an experimental population have been determined, QTL identification can be accomplished. We describe the statistical tools used to identify the existence, location, and significance of QTLs. These different methods have advantages and disadvantages to consider when selecting the appropriate model to be used, which is briefly discussed.Although the objective of QTL mapping is to identify genomic regions associated with a trait, the ultimate goal is to identify the gene and the genetic variation (which is often quantitative trait nucleotide, QTN) or haplotype that is responsible for the phenotype. By discovering the function of causative variants or haplotypes we can understand the molecular changes that lead to the phenotype. We briefly describe how the genomic sequences can be exploited to identify QTNs and how these can be validated in congenic strains and functionally tested to understand their influence on phenotype expression.