Random rapid amplification of cDNA ends (RRACE) allows for cloning of multiple novel human cDNA fragments containing (CAG)n repeats.
We describe a new technique for isolating cDNA fragments in which (i) either a partial sequence of the cDNA is known or (ii) 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.
Carney
J P
JP
Department of Pediatrics, Riley Hospital for Children, Indiana University Medical Center, Indianapolis 46202, USA.
McKnight
C
C
VanEpps
S
S
Kelley
M R
MR
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification*; D005784:Gene Amplification; D006801:Humans; D015458:Leukemia, T-Cell / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D012313:RNA / Q000032:analysis; D012091:Repetitive Sequences, Nucleic Acid; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D012313:RNA