Fast Slab Gel Electrophoretic Separation of DNA Fragments with a Short Migration Distance Using Thermal Lens Microscope.

Abstract

We demonstrated fast separation of DNA fragments in a short slab gel through the minimization of band-broadening. A discontinuous buffer system was used to reduce band-broadening from sample introduction, and a high spatial resolution thermal lens microscope (TLM) was used to reduce such an effect from the detector. Bands of DNA fragments were sharpened into 100 m m, which was about 1/10 of the bandwidths in conventional continuous slab gel electrophoresis. The 100 base pair ladder DNA sample, which ranges from 100 bp to 2000 bp, was completely separated within 15 min with a migration distance of 18 mm and was detected by TLM after silver staining. These results suggested the possibility of fast separation of DNA fragments in a miniaturized discontinuous slab gel. As well as the inherent properties of parallel separation of slab gel, high throughput separation can be obtained, which is significant for DNA analyses such as DNA sequencing, genetic diagnosis and forensic identification analyses.