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 Kelley

Research output: Contribution to journalArticle

9 Citations (Scopus)

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

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Organism Cloning
Complementary DNA
Trinucleotide Repeats
Clone Cells
Genes
Expressed Sequence Tags
Nucleic Acid Databases

Keywords

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

ASJC Scopus subject areas

  • Genetics

Cite this

Random rapid amplification of cDNA ends (RRACE) allows for cloning of multiple novel human cDNA fragments containing (CAG)n repeats. / Carney, James P.; McKnight, Christopher; Vanepps, Steffany; Kelley, Mark.

In: Gene, Vol. 155, No. 2, 03.04.1995, p. 289-292.

Research output: Contribution to journalArticle

Carney, James P. ; McKnight, Christopher ; Vanepps, Steffany ; Kelley, Mark. / Random rapid amplification of cDNA ends (RRACE) allows for cloning of multiple novel human cDNA fragments containing (CAG)n repeats. In: Gene. 1995 ; Vol. 155, No. 2. pp. 289-292.
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