Titration of variant DNA sequences differing by a single point-mutation by selective dot-blot hybridization with synthetic oligonucleotides |
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Authors: | M Nicoloso H Q Liang J P Bachellerie |
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Institution: | Centre de Recherches de Biochimie et Génétique Cellulaires du CNRS, Université Paul-Sabatier, Toulouse, France. |
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Abstract: | A dot-blot hybridization procedure with synthetic oligonucleotide probes is reported, which allows the quantitative titration in genomic DNA of variant forms of repeated genes differing by a single nucleotide change. It involves the utilization of a pair of 22-base long oligonucleotides matching the two variant sequences and the choice of an hybridization temperature very close to the Td of the oligonucleotide/DNA duplexes. The selectivity is achieved through a competition between the cognate labeled and the non-cognate unlabeled probes in the hybridization mixture. |
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