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Detection of β‐thalassaemia mutations using DNA heteroduplex generator molecules
Author(s) -
SAVAGE DAVID A.,
WOOD NIGEL A. P.,
BIDWELL JEFFREY L.,
FITCHES ALISON,
OLD JOHN M.,
HUI KAM M.
Publication year - 1995
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1995.tb05585.x
Subject(s) - heteroduplex , polymerase chain reaction , mutation , dna , population , genetics , microbiology and biotechnology , biology , mutant , gene , medicine , environmental health
Summary. In this report we describe a rapid polymerase chain reaction (PCR) based method for the detection of β‐thalassaemia (β‐thal) mutations. This method is based on the visualization of unique DNA heteroduplex banding patterns, following non‐denaturing poly aery lamide gel electrophoresis, resulting from hybridization between mutant PCR products and synthetic DNA heteroduplex generator molecules. Using the Singaporean population, which consists of Chinese, Malay and Asian Indian ethnic groups, as a model, we have constructed and evaluated three DNA heteroduplex generator molecules for the detection of the common β‐thalassaemia mutations found in this population. The results show that these three molecules are capable of detecting approximately 95% of the mutations found in the Singaporean population. We propose that this technology may be applied as an alternative screening strategy for β‐thalassaemia mutations because it is technically simple, flexible, cost‐effective, and requires only minimal laboratory resources.