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Detection of the DeltaF508 trinucleotide deletion in the human CFTR gene by UV immobilization on nylon of a complex of a PCR fragment with a highly specific probe
Authors:Kabilov M R  Pyshny? D V  Dymshits G M  Zarytova V F  Ivanova E M
Institution:Novosiberian Institute of Bioorganic Chemistry, Institute of Cytology and Genetics, Siberian Division, Russian Academy of Sciences, Novosibirsk, 630090 Russia.
Abstract:Deletion delta F508 has been revealed in PCR-amplified regions of human gene CFTR by color detection of the hybridization complex obtained by ligation of a tandem of short oligonucleotides on a DNA template followed by UV immobilization on nylon. The method allows reliable detection of the three-nucleotide deletion (insertion). The nonspecific signal depends on the nucleotide composition of the biotinylated tandem component. A significant level of the specific signal was achieved by using the PCR-amplified DNA fragments of different length (200-400 bp) irrespective of the position of the tandem-binding site in their sequences.
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