Miniaturized Sequencing Gel System for Quick Analysis of DNA by Hydroxyl Radical Cleavage |
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Authors: | Taichi E Takasuka Yi-Ju Hsieh Arnold Stein |
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Institution: | 1. Department of Biochemistry, University of Wisconsin-Madison, Madison, WI, USA 3. Department of Biological Sciences, Purdue University, West Lafayette, IN, USA 2. Department of Urology, Stanford University, Stanford, CA, USA
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Abstract: | We described a simple and quick miniaturized sequencing gel system for DNA analysis. Two major modifications were made to the previously reported miniaturized DNA sequencing gel system to achieve high-resolution hydroxyl radical cleavage analysis: including formamide in the miniaturized gel and providing uniform heating during electrophoresis. Our method enables one to reduce the cost for chemicals and to significantly reduce electrophoresis time. Furthermore, minimal gel handling simplifies the entire process. We show that the resolution of DNA fragments obtained by hydroxyl radical cleavage for the miniaturized gel is similar to that of a large conventional sequencing gel. |
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