A new approach to revealing point mutations in DNA analyzed by colorimetric detection.
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.
Kabilov
Marsel
M
Institute of Chemical Biology and Fundamental Medicine, SD, RAS, Novosibirsk, Russia. kmr@yandex.ru
Pyshnyi
Dmitrii
D
Dymshits
Grigorii
G
Zarytova
Valentina
V
Ivanova
Evgenia
E
D001483:Base Sequence; D003124:Colorimetry / Q000379:methods*; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics; D009693:Nucleic Acid Hybridization; D017354:Point Mutation; D014466:Ultraviolet Rays
D017931:DNA Primers; D004279:DNA, Viral