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 |
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Authors: | Kabilov M R Pyshny? D V Dymshits G M Zarytova V F Ivanova E M |
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Institution: | Novosiberian Institute of Bioorganic Chemistry, Institute of Cytology and Genetics, Siberian Division, Russian Academy of Sciences, Novosibirsk, 630090 Russia. |
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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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