Isolation of molecular markers from specific chromosomal intervals using DNA pools from existing mapping populations

James J. Giovannoni, Rod A Wing, Martin W. Ganal, Steven D. Tanksley

Research output: Contribution to journalArticle

233 Citations (Scopus)

Abstract

We present a general method for isolating molecular markers specific to any region of a chromosome using existing mapping populations. Two pools of DNA from individuals homozygous for opposing alleles for a targeted chromosomal interval, defined by two or more linked RFLP markers, are constructed from members of an existing mapping population. The DNA pools are then screened for polymorphism using random oligonuclotide primers and PCR (1). Polymorphic DNA bands should represent DNA sequences within or adjacent to the selected interval. We tested this method in tomato using two genomic intervals containing genes responsible for regulating pedicle abscission pointless) and fruit ripening (non-ripening). DNA pools containing 7 to 14 F2 individuals for each interval were screened with 200 random primers. Three polymorphic markers were thus identified, two of which were subsequently shown to be tightly linked to the selected intervals. The third marker mapped to the same chromosome (11) but 45 cM away from the selected interval. A particularly attractive attribute of this method is that a single mapping population can be used to target any interval in the genome. Although this method has been demonstrated in tomato, it should be applicable to any sexually reproducing organism for which segregating populations are being used to construct genetic linkage maps.

Original languageEnglish (US)
Pages (from-to)6553-6558
Number of pages6
JournalNucleic Acids Research
Volume19
Issue number23
StatePublished - Dec 11 1991
Externally publishedYes

Fingerprint

Isolation
DNA
Interval
Lycopersicon esculentum
Chromosomes
Population
Genes
Tomato
Chromosomes, Human, Pair 11
Genetic Linkage
DNA sequences
Chromosome
Fruits
Polymorphism
Restriction Fragment Length Polymorphisms
Fruit
Abscission
Alleles
Genome
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Isolation of molecular markers from specific chromosomal intervals using DNA pools from existing mapping populations. / Giovannoni, James J.; Wing, Rod A; Ganal, Martin W.; Tanksley, Steven D.

In: Nucleic Acids Research, Vol. 19, No. 23, 11.12.1991, p. 6553-6558.

Research output: Contribution to journalArticle

Giovannoni, James J. ; Wing, Rod A ; Ganal, Martin W. ; Tanksley, Steven D. / Isolation of molecular markers from specific chromosomal intervals using DNA pools from existing mapping populations. In: Nucleic Acids Research. 1991 ; Vol. 19, No. 23. pp. 6553-6558.
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