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Nucleic Acids Research, 1995, Vol. 23, No. 21 4415-4420
© 1995

Articles

Direct selection of cDNAs using whole chromosomes

Sylvie Rouquier1, Barbara J. Trask2, Sylvie Taviaux1, Ger van den Engh2, Sylvie Diriong1, Gregory G. Lennon3 and Dominique Giorg1,*

1Centre de Recherche de Biochimie Macromol`culaire CNRS UPR 9008, BP 5051,1919 Route de mende, 34033 Montpellier cedex, France 2Department of Molecular Biotechnology FJ-20, University of Washington Seattle, WA 98195, USA 3Lawrence Livermore National Laboratory PO Box 808, L-452, Livermore,CA 94551, USA

*To whom correspondence should be addressed

Received July 10, 1995. Accepted September 25, 1995.

We have developed a method for direct selection of cDNAs using whole chromosomes as target DNA. Double-strand cDNAs were synthesized from human fetal brain polyadenylated mRNAs. Flow-sorted chromosomes 17 and 19 were amplified by degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) and used to capture ds cDNAs by an improved magnetic bead capture protocol. To demonstrate the capabilities of this method, the selected cDNAs were used as probes in FISH experiments. The selected cDNA populations specifically painted chromosomes 17 or 19 on metaphase spreads. These results demonstrate that it is possible to do chromosome painting using cDNA probes and that this method is a means to rapidly select expressed sequences encoded by any portion of the genome


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