Alu-splice PCR: A simple method to isolate exon-containing fragments from cloned human genomic DNA

Juan José Fuentes, Carles Pucharcós, Melanie Pritchard, Xavier P. Estivill

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6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)346-350
Number of pages5
JournalHuman Genetics
Volume101
Issue number3
DOIs
Publication statusPublished - 1997
Externally publishedYes

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ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

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