Production of monoclonal anti-thymine-dimer antibody and its use for detection of STR alleles.
To simplify the manual detection of STRs, we developed an immunoblotting method using an anti-thymine-dimer antibody. Upon ultraviolet irradiation, adjacent thymine residues in a DNA strand readily form thymine dimers which are detectable by anti-thymine-dimer antibody. We simply irradiated the supernatant of a boiled suspension of P3U1 myeloma cells with ultraviolet rays, and immunized BALB/c mice with it. By fusing their popliteal lymph node cells with the P3U1 myeloma cell line, we produced a monoclonal anti-thymine-dimer antibody. Using this antibody, we were able to detect STR alleles containing AAAG, AAGG, or AATG repeats by immunoblotting. Our method is well suited for use in parentage testing.
Fukuda
M
M
Department of Forensic Medicine, Oita Medical University, Japan.
Tamaki
Y
Y
Kishida
T
T
Wang
W
W
D000483:Alleles; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001483:Base Sequence; D015151:Immunoblotting; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011740:Pyrimidine Dimers / Q000276:immunology*; D012091:Repetitive Sequences, Nucleic Acid; D014466:Ultraviolet Rays
D000911:Antibodies, Monoclonal; D011740:Pyrimidine Dimers