Analytical Sciences

Abstract − Analytical Sciences, 15(3), 223 (1999).

Fast Slab Gel Electrophoretic Separation of DNAFragments with a Short Migration Distance Using Thermal Lens Microscope
Jinjian ZHENG*, TamaoODAKE**, Takehiko KITAMORI* , ** and Tsuguo SAWADA* *
*Department of Applied Chemistry, School of Engineering, The University of Tokyo, Hongo, Bunkyo, Tokyo 113-8656, Japan
**Kanagawa Academy of Science and Technology, 3-2-1 Sakado, Takatsu,Kanagawa 213-0012, Japan
We demonstrated fast separation of DNA fragments ina short slab gel through the minimization of band-broadening. Adiscontinuous buffer system was used to reduce band-broadening from sampleintroduction, and a high spatial resolution thermal lens microscope (TLM)was used to reduce such an effect from the detector. Bands of DNA fragmentswere sharpened into 100 mm, which was about 1/10 of the bandwidths inconventional continuous slab gel electrophoresis. The 100 base pair ladderDNA sample, which ranges from 100 bp to 2000 bp, was completely separatedwithin 15 min with a migration distance of 18 mm and was detected by TLMafter silver staining. These results suggested the possibility of fastseparation of DNA fragments in a miniaturized discontinuous slab gel. Aswell as the inherent properties of parallel separation of slab gel, highthroughput separation can be obtained, which is significant for DNAanalyses such as DNA sequencing, genetic diagnosis and forensicidentification analyses. (Keywords: Electrophoresis, slab gel, DNA, miniaturization, thermallens microscope, discontinuous buffer)