Titration of variant DNA sequences differing by a single point-mutation by selective dot-blot hybridization with synthetic oligonucleotides.
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.
Nicoloso
M
M
Centre de Recherches de Biochimie et Génétique Cellulaires du CNRS, Université Paul-Sabatier, Toulouse, France.
Liang
H Q
HQ
Bachellerie
J P
JP
D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D005796:Genes; D014644:Genetic Variation; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal / Q000235:genetics*; D012339:RNA, Ribosomal, 28S / Q000235:genetics*; D015183:Restriction Mapping; D013816:Thermodynamics
D015345:Oligonucleotide Probes; D012335:RNA, Ribosomal; D012339:RNA, Ribosomal, 28S; D004247:DNA