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- W2113296885 abstract "We have read with interest the recent reports of Marziliano et al. (1) and Pirulli et al. (2), who used melting temperature assays to genotype different types of mutations. Because detection of the underlying mutation is only indirect in these methods, they differ in their molecular detection principle from hybridization probe-based genotyping (3) or allele-specific amplification coupled to SYBR-green I detection (4). Indirect detection methods demand extra caution in the assignment of genotypes solely on the basis of product melting temperature ( T m). From a theoretical standpoint, some genotypes will not be detected.The UDP-glucuronosyltransferase 1 (UGT1A1) (TA)n insertion/deletion polymorphism is of significance for the manifestation of Gilbert disease (3)(5). TA repeats are intrinsically unstable, and, therefore, it is not surprising that five to eight TA repeats occur in humans and have functional significance (5). The method of Marziliano et al. (1) uses T m as a measure of amplicon length to discover the UGT1A1 promoter genotype. In the described assay, a 2-bp difference is detected by the resulting T m shift in a 130-bp amplicon. A similar technique was successfully used in the screening for a 9-bp deletion in a 55-bp PCR amplicon (6). However, for the assay of Marziliano et al. (1), it has to be noted that using …" @default.
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- W2113296885 date "2001-07-01" @default.
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- W2113296885 title "Limitations of Genotyping Based on Amplicon Melting Temperature" @default.
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- W2113296885 doi "https://doi.org/10.1093/clinchem/47.7.1331" @default.
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