Trost P, Guttman A
Genetic BioSytems, Inc., San Diego, California 92121, USA.
Anal Chem. 1998 Sep 15;70(18):3930-5. doi: 10.1021/ac980359u.
High-throughput DNA sequencing techniques are under rapid development currently, mainly triggered by the Human Genome Project. At the present time, slab gel based automated DNA sequencing is the standard procedure, utilizing fluorophore labeling and laser-induced fluorescence detection with scanning technology. In this paper, a novel, fiber-optic bundle based detection system is introduced, where a central illuminating fiber is used for the excitation of the electrophoretically separated fluorophore-labeled DNA sequencing fragments, along with several collecting fibers disposed around the illuminating fiber to collect the emitted fluorescent signal. As a model system, Cy5-labeled DNA sequencing fragments were separated on an ultrathin polyacrylamide slab gel and detected by the fiber bundle based laser-induced fluorescence detection system. A 640-nm diode laser was used to generate the illumination beam, and the emitted light collected by the fiber bundle was detected by a solid-state avalanche photodiode.
高通量DNA测序技术目前正在迅速发展,主要由人类基因组计划推动。目前,基于平板凝胶的自动化DNA测序是标准程序,利用荧光团标记和激光诱导荧光检测及扫描技术。本文介绍了一种新型的基于光纤束的检测系统,其中一根中央照明光纤用于激发经电泳分离的荧光团标记的DNA测序片段,同时在照明光纤周围布置几根收集光纤以收集发射的荧光信号。作为一个模型系统,Cy5标记的DNA测序片段在超薄聚丙烯酰胺平板凝胶上进行分离,并通过基于光纤束的激光诱导荧光检测系统进行检测。使用640纳米二极管激光器产生照明光束,由光纤束收集的发射光由固态雪崩光电二极管进行检测。