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

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 oligonucleotide 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 (jointless) 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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Giovannoni JJ,Wing RA,Ganal MW,Tanksley SD

doi

10.1093/nar/19.23.6553

subject

Has Abstract

pub_date

1991-12-11 00:00:00

pages

6553-8

issue

23

eissn

0305-1048

issn

1362-4962

journal_volume

19

pub_type

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