A robust method for detecting CHK2/RAD53 mutations in genomic DNA

Nayanta Sodha, Richard S. Houlston, Richard Williams, Martin A. Yuille, John Mangion, Rosalind A. Eeles

    Research output: Contribution to journalArticlepeer-review


    While screening for germline CHK2 mutations in cancer cases by heteroduplex CSGE, we observed that additional PCR fragments were generated from the 3′ end region of the gene that includes exons 11-14. Direct sequencing of these fragments suggested that homologous loci (possibly pseudogenes) were concomitantly being amplified. Searches of public sequence databases showed that a number of areas of the genome show a high degree of homology to exons 10-14 of the CHK2 gene. The presence of these homologous regions means that standard screening methods for detecting mutations in CHK2, based on PCR of genomic DNA, are prone to error. To circumvent this problem, we have developed a strategy, based on long-range PCR, to screen the functional copy of CHK2. Using this approach it is possible to carry out a comprehensive mutational analysis of CHK2 from genomic DNA. © 2002 Wiley-Liss, Inc.
    Original languageEnglish
    Pages (from-to)173-177
    Number of pages4
    JournalHuman Mutation
    Issue number2
    Publication statusPublished - 2002


    • CHK2
    • CSGE
    • Homologous loci
    • Long range PCR
    • Mutation analysis
    • Pseudogene interference
    • RAD53


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