Chromosomal assignment of human YAC clones by fluorescence in Situ hybridization: Use of single-yeast-colony PCR and multiple labeling

Antonio Baldini, Mark Ross, Dean Nizetic, Radost Vatcheva, Elizabeth A. Lindsay, Hans Lehrach, Marcello Siniscalco

Research output: Contribution to journalArticlepeer-review

Abstract

Alu-PCR provides a convenient tool for amplification of human-specific sequences from yeast DNA containing yeast artificial chromosomes (YAC) clones. PCR products can be labeled nonisotopically and hybridized in situ, and the chromosomal origin of the clones can be determined. This avoids time-consuming gel purification of the yeast artificial chromosome and the low-efficiency procedure of labeling whole yeast DNA containing the YAC. The application of Alu-PCR to single-yeast colonies permits the mapping of YACs at a very early stage of their characterization. In situ hybridization can detect clones with noncontiguous fragments of DNA, and these can be discarded without further time-consuming characterization. To increase further the potential of the method, we show the application of multicolor hybridization techniques.

Original languageEnglish
Pages (from-to)181-184
Number of pages4
JournalGenomics
Volume14
Issue number1
DOIs
Publication statusPublished - 1992

ASJC Scopus subject areas

  • Genetics

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