A new approach to revealing point mutations in DNA analyzed by colorimetric detection

作者：Kabilov M^*; Pyshnyi D; Dymshits G; Zarytova V; Ivanova E

来源：Nucleosides Nucleotides & Nucleic Acids, 2004, 23(6-7): 1023-1030.

DOI：10.1081/NCN-200026060

• T4 DNA ligase
• ligation
• hybridization
• high-specific probe
• colorimetric detection
• point mutation
• HIV
• Delta F508

A new approach to detection of point mutations in an amplified DNA was developed. The approach is based on highly selective ligation (T4 DNA ligase) of a tandem of short oligonucleotides one of which contains the biotin group. The ligation product is formed only when the hybridization complex DNA/tandem is formed and the tandem is perfect. The hybridization complex DNA/(biotinylated ligation product) was separated from the biotinylated component of the tandem by UV-immobilization of the reaction mixture on a nylon membrane. The immobilized hybridization complex was detected colorimetrically by a streptavidin-alkaline phosphatase cojugate with a chromogenic substrate.

• 出版日期2004-10