An oligonucleotide-tagged microarray for routine diagnostics of colon cancer by genotyping KRAS mutations.

作者: YULIANG LIU , HAUKUR GUDNASON , YI-PING LI , DANG DUONG BANG , ANDERS WOLFF

DOI: 10.3892/IJO.2014.2541

关键词:

摘要: Colorectal cancer (CRC) is one of the most prevalent types cancer, causing significant morbidity and mortality worldwide. CRC curable if diagnosed at an early stage. Mutations in oncogene KRAS play a critical role development CRC. Detection activated diagnostic therapeutic importance. In this study, gene fragments containing mutations codon 12 were amplified by multiplex PCR using 5'-Cy5-labeled reverse primer combination with 3'-mutation-specific forward primers that linked four unique nucleotide-sequence tags 5'-end. The Cy5-labeled was extended under amplification to 5'-end mutation-specific thus included complimentary sequence tag. products hybridized tag-probes immobilized on various substrates detected scanner. Our results indicate all derived from cells clinical samples could be unambiguously detected. accurately when mutant DNA present only 10% starting mixed materials including wild-type genomic DNA, which isolated either or spiked fecal samples. stable multiple thermal cycling treatments, allowing re-use tag-microarray further optimization solid PCR. demonstrated novel oligonucleotide-tagged microarray system has been developed would suitable used for detection diagnosis

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