Pulsed-field gel electrophoresis is a frequent technique used to separate exceptionally large DNA fragments. In a typical continuous field electrophoresis, it is challenging to separate DNA fragments larger than 20kbp because they migrate at a comparable rate. To overcome this challenge, it is necessary to develop a novel matrix for the electrophoresis. Here, we describe the electrophoresis of large DNA fragments up to 166kbp using a supramolecular gel matrix and a typical continuous field electrophoresis system. C-3-symmetric tris-urea self-assembled into a supramolecular hydrogel in tris-boric acid-EDTA buffer, a typical buffer for DNA electrophoresis, and the supramolecular hydrogel was used as a matrix for electrophoresis to separate large DNA fragments. Three types of DNA marker, the -Hind III digest (2 to 23kbp), Lambda DNA-Mono Cut Mix (10 to 49kbp), and Marker 7 GT (10 to 165kbp), were analyzed in this study. Large DNA fragments of greater than 100kbp showed distinct mobility using a typical continuous field electrophoresis system.