[Screening C282Y mutation with double-stranded probes using synchronous fluorometry].

Yi Chuan

The Key Laboratory of Cell Biology and Tumor Cell Engineering of The Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, Fujian 361005, China.

Published: January 2003

We described in the paper a new high-throughput screening method for Cys282Tyr mutation in hereditary haemochromatosis with double-stranded probe using synchronous fluorometry. The probe for wild type was labeled with Fam, the probe for mutant type was labeled with Joe. After PCR, reaction tubes were transferred to a spectrofluorometer, where synchronous spectra were scanned in a constant-wavelength mode. The genotype could be obtained through the appearance of the fluorescence peaks corresponding to each probe. The results were totally in agreement with restriction endonuclease analysis. Considering the simplicity,low cost and specificity, this approach could be generally applied to detect varieties of gene mutations.

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