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

Research output: Contribution to journalArticle

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

Fingerprint

Alu Elements
Exons
Polymerase Chain Reaction
DNA
Chromosomes, Human, Pair 21
RNA Splice Sites
Consensus Sequence
Human Chromosomes
Sequence Analysis
Clone Cells
Genome

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

Cite this

Alu-splice PCR : A simple method to isolate exon-containing fragments from cloned human genomic DNA. / Fuentes, Juan José; Pucharcós, Carles; Pritchard, Melanie; Estivill, Xavier P.

In: Human Genetics, Vol. 101, No. 3, 1997, p. 346-350.

Research output: Contribution to journalArticle

Fuentes, Juan José ; Pucharcós, Carles ; Pritchard, Melanie ; Estivill, Xavier P. / Alu-splice PCR : A simple method to isolate exon-containing fragments from cloned human genomic DNA. In: Human Genetics. 1997 ; Vol. 101, No. 3. pp. 346-350.
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