Rapid and large-scale method to detect K-ras gene mutations in tumor samples.
We have developed a rapid and large-scale method for the detection of K-ras gene mutations in tumors. First, DNA is amplified by an asymmetric PCR; second, the single-strand dinitrophenyl (DNP)-labeled amplified DNA is hybridized specifically to oligonucleotide probes affixed on a tube. Finally, perfectly matched duplexes are easily detected by a monoclonal anti-DNP antibody bearing 125I. The usefulness of this technique is illustrated by analyzing K-ras codon 12 mutations in human colorectal samples. This reliable assay procedure can be applied to the rapid screening of virtually any genetic disease caused by previously described point mutations.
Lopez-Crapez
E
E
C.R.L.C. Val d'Aurelle, Laboratoire de Radioanalyse, Montpellier, France.
Chypre
C
C
Saavedra
J
J
Marchand
J
J
Grenier
J
J
D000368:Aged; D001706:Biopsy; D003062:Codon; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D015342:DNA Probes; D004273:DNA, Neoplasm / Q000032:analysis* / Q000235:genetics; D005260:Female; D011905:Genes, ras; D015894:Genome, Human; D006801:Humans; D008297:Male; D008875:Middle Aged; D009693:Nucleic Acid Hybridization; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods
D003062:Codon; D017931:DNA Primers; D015342:DNA Probes; D004273:DNA, Neoplasm