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Positional Cloning


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Positional cloning, also known as map-based cloning, is a technique for the positioning of a trait-associated gene in the genome and involves methods such as linkage analysis, association mapping, and bioinformatics. Due to its high efficiency and accuracy, the positional cloning technique has shed light on modern plant breeding for the identification of genes or quantitative trait loci (QTLs) associated desired traits, and can potentially accelerate marker-assisted selection as well as introduction of valuable traits. Lifeasible, as a leading company in plant breeding technologies, offers tailored positional cloning service for our customers worldwide.

The principle of positional cloning is to isolate a particular allele from a cDNA library with specific primers, using chromosome waking or thermal asymmetric interlaced PCR (tail-PCR), which are used in the identification of unknown regions flanking a known DNA sequence. While cloning of QTLs requires information of both genetic and physical maps, and is performed following a series of steps (Figure 1):

  • Identification of linkage group: Both gynogenic half-tetrad diploids and bulked segregant analysis (BSA) can be used.
  • Construction of genetic map.
  • Conversion of the genetic map to a physical map.
  • Construction of fine-scale physical map.
  • Candidate gene selection.
  • Validation of candidate QTLs function.


Figure 1.  Flow chart for positional cloning in rice (Koh et al., 2015).

At Lifeasible, our teams of geneticists, statisticians, and software engineers will work closely toward the most optimized solvent for any given project, providing both scientific and technical insights for population type, sample size, marker collection, and other critical aspects of the positional cloning program, to ensure the success of your projects. Welcome to contact us for technical consulting, experimental design, and other informations. We are always happy to walk you through our featured technologies, service plans, collaboration options, and more.

Reference:

  1. Koh, H. J., Kwon, S. Y., and Thomson, M. (2015). Current technologies in plant molecular breeding. Springer Dordrecht Heidelberg, New York, London.
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