Highly sensitive and direct detection of DNA fragments using a laser-induced capillary vibration effect.
A pulsed laser-induced stationary wave capillary vibration detection method was applied to the sensitive detection of capillary gel electrophoresis, and the direct detection of non-labeled nucleic acids, such as DNA sequencing products, was demonstrated. An excimer laser operating at 248 nm was used as a CVL excitation source, and polynucleotides were sensitively detected without derivatization. From an investigation on the endurance of several matrixes to pulsed laser irradiation, a polyacrylamide without a cross-linker (0%C) was found to have adequate endurance, and it exhibited no serious damage during an analysis. A cytosine-terminated sequence reaction product was detected with a sensitivity close to that of laser-induced fluorometry (LIF). These results suggest the feasibility of the highly sensitive detection of ultramicro amounts of biological materials without a pre- or post-column derivatization, which has usually been required in sensitive detection procedures, such as LIF. Furthermore, the feasibility of a novel DNA sequencing method is also suggested.
Odake
T
T
Department of Chemistry, Gunma University, Kiryu, Japan.
Tsunoda
K
K
Kitamori
T
T
Sawada
T
T
D000180:Acrylic Resins / Q000528:radiation effects; D004247:DNA / Q000737:chemistry* / Q000528:radiation effects; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D007834:Lasers; D011061:Poly A / Q000737:chemistry; D017422:Sequence Analysis, DNA; D014732:Vibration; D000099166:Polyacrylamides
D000180:Acrylic Resins; D004247:DNA; D011061:Poly A; D000099166:Polyacrylamides