Mutation detection in K-ras, BRCA1, BRCA2, and p53 using PCR/LDR and a universal DNA microarray.
We have developed a multiplex PCR/ligase detection reaction (PCR/LDR) that combines high sensitivity with the ability to simultaneously detect hundreds of mutations in a single-tube reaction. To enable us to rapidly assay large numbers of samples, we have linked this mutation detection scheme with analysis on a Universal DNA microarray. We have successfully applied this approach to characterize K-ras and p53 mutations in DNA derived from undissected colon tumors. The sensitivity of the assay has also facilitated detection of low-frequency mutations in BRCA1 and BRCA2 in pooled samples of DNA; thus, PCR/LDR can rapidly screen large numbers of DNA samples required for population studies.
Favis
R
R
Department of Microbiology, Hearst Microbiology Research Center, Joan and Sanford I. Weill Medical College of Cornell University, New York, New York 10021, USA.
Barany
F
F
D004247:DNA / Q000235:genetics*; D011088:DNA Ligases; D016147:Genes, Tumor Suppressor; D011905:Genes, ras; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*
D004247:DNA; D011088:DNA Ligases