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Background: To compare high resolution melting (HRM) and primer extension (PE) for mutation screening of KRAS codons 12 and 13.
Methods: DNA samples were isolated from 60 colorectal cancer (CRC) patients. HRM and primer extension analyses were used for mutation screening of KRAS codons 12 and 13.
Results: Both methods can detect KRAS mutations in a DNA mixture containing as little as 5 % mutant DNA. The concordant rate between the two methods was 100 %. HRM analysis is able to distinguish mutant from wild type, however, it is unable to detect the actual base change. PE analysis needs more procedures, is time-consuming and slightly more expensive but it is able to detect the precise mutation.
Conclusions: This study showed that HRM is a reliable screening method to identify mutations in the KRAS gene, and the PE can be used as a diagnostic method to accurately pinpoint the nature of mutations.
DOI: Clin. Lab. 2010;56:175-186
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