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Nucleic Acids Research, 1994, Vol. 22, No. 22 4806-4809
© 1994

MOLECULAR BIOLOGY

PCR-based immortalization and screening of hierarchical pools of cDNAs

Maurizio D'Esposito1,2, Richard Mazzarella1, Gina Pengue1, Carmela Johns1, Michele D'Urso2 and David Schlessinger1,*

1Department of Molecular Microbiology and Center for Genetics in Medicine, Washington University Medical School St Louis, MO 63110, USA 2International Institute of Genetics and Biophysics CNR, Naples 80125, Italy

*To whom correpsondence should be addressed

Received May 28, 1994. Revised August 23, 1994. Accepted August 23, 1994.

Starting from sequences of at least 60 bp, PCR-based screening has been developed to recover cDNAs from libraries without the necessity for hybridization or extensive DNA extraction steps. The method maintains the indefinite availability of even scarce cDNA libraries and provides an estimate of the relative abundance of the mRNA species. Isolation of a cDNA clone can be done in less than a week. cDNAs were isolated that were cognate for fragments of expressed sequences and for an exon predicted from genomic sequence.


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