Random rapid amplification of cDNA ends (RRACE) allows for cloning of multiple novel human cDNA fragments containing (CAG)n repeats

James P. Carney, Christopher McKnight, Steffany Vanepps, Mark R. Kelley

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

We describe a new technique for isolating cDNA fragments in which (i) either a partial sequence of the cDNA is known or (i) a repeat sequence is utilized. We have used this technique, termed random rapid amplification of cDNA ends (random RACE), to isolate a number of trinucleotide repeat (CAG)n-containing genes. Using the random RACE (RRACE) technique, we have isolated over a hundred (CAG)n-containing genes. The results of our initial analysis of ten clones indicate that three are identical to previously cloned (CAG)n-containing genes. Three of our clones matched with expressed sequence tags, one of which contained a CA repeat. The remaining four clones did not match with any sequence in GenBank. These results indicate that this approach provides a rapid and efficient method for isolating trinucleotide repeat-containing cDNA fragments. Finally, this technique may be used for purposes other than cloning repeat-containing cDNA fragments. If only a partial sequence of a gene is known, our system, described here, provides a rapid and efficient method for isolating a fragment of the gene of interest.

Original languageEnglish (US)
Pages (from-to)289-292
Number of pages4
JournalGene
Volume155
Issue number2
DOIs
StatePublished - Apr 3 1995

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Keywords

  • CAG repeat gene
  • genetic disorder
  • PCR cloning
  • Trinucleotide repeat expansion

ASJC Scopus subject areas

  • Genetics

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