Detection of single-nucleotide polymorphisms in cancer-related genes by minisequencing on a microelectronic DNA chip.
The ability to realize simultaneous genotyping of multiple single-nucleotide polymorphisms or mutations is valuable in DNA samples from complex multigenic pathologies such as cancer. In this way, the complexity (number of hybridization units per chip) of the developed MICAM DNA chip, and the orientation of the grafted pyrrole oligonucleotides, make it particularly well adapted to the analysis of single-nucleotide polymorphisms/mutations in multiple potential tumoral markers. The proposed genotyping methodology is based on solid-phase minisequencing, where oligonucleotides are designed to anneal immediately upstream of the polymorphism sites, and labeled dideoxynucleotides are used as substrates for polymerase extension. The developed assay was applied to the analysis of the TP53 codon 72 polymorphism on DNA from cell lines and human colorectal samples.
Ho-Pun-Cheung
Alexandre
A
Centre de Recherche en Cancérologie, Centre Régional de Lutte contre le Cancer Val d'Aurelle-Paul Lamarque, Parc Euromédecine, Montpellier Cedex, France.
Abaibou
Hafid
H
Cleuziat
Philippe
P
Lopez-Crapez
Evelyne
E
D045744:Cell Line, Tumor; D015342:DNA Probes; D004273:DNA, Neoplasm / Q000302:isolation & purification; D004581:Electronics / Q000295:instrumentation*; D005453:Fluorescence; D052138:Genes, Neoplasm; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D009691:Nucleic Acid Denaturation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009841:Oligonucleotides; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*
D015342:DNA Probes; D004273:DNA, Neoplasm; D009841:Oligonucleotides