Abstract
We have developed a simple, straightforward procedure to isolate exons from cloned human genomic DNA. The method is PCR based and relies upon the conservation of splice-site sequences and the frequency of Alu repeat elements in the genome to capture coding sequences. We designed two different sets of primers: a primer from each end of the Alu element and primers with the 5′ or 3′ splice-site consensus sequences. Putative exons were amplified by PCR using YAC DNA as starting material. We applied Alu-splice PCR to two overlapping YACs, 72H9 and 860G11, from human chromosome 21. Sequence and northern analysis of 37 initial clones resulted in the identification of five novel exons.
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Received: 17 July 1997 / Accepted: 28 August 1997
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Fuentes, JJ., Pucharcós, C., Pritchard, M. et al. Alu-splice PCR: a simple method to isolate exon-containing fragments from cloned human genomic DNA. Hum Genet 101, 346–350 (1997). https://doi.org/10.1007/s004390050639
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DOI: https://doi.org/10.1007/s004390050639