High-throughput microsatellite analysis using fluorescent dUTPs for high-resolution genetic mapping of the mouse genome.

  1. M Rhodes,
  2. A Dearlove,
  3. R Straw,
  4. S Fernando,
  5. A Evans,
  6. M Greener,
  7. T Lacey,
  8. M Kelly,
  9. K Gibson,
  10. S D Brown, and
  11. C Mundy
  1. MRC Human Genome Mapping Project Resource Centre, Hinxton, Cambridge, UK.

Abstract

The use of fluorescent end-labeled primers has proved successful for rapid, semiautomated genotyping of microsatellite loci. However, custom synthesis is expensive and costs can be prohibitive when a wide range of markers is to be analyzed for only a few genotypings. This particularly applies to high-resolution genetic mapping in the mouse either in the construction of global maps or in the production of local high-resolution genetic maps for positional cloning. We demonstrate here the use of fluorescent dUTPs for cost-effective, high-throughput microsatellite genotyping in the mouse. This alternative to the use of fluorescent end-labeled primers for semiautomated genotyping is potentially applicable to the construction of linkage maps in other species.

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