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Soybean is one of the important commodities in Indonesia, but its production is still low due to the attack of the fungus Athelia rolfsii Curzi. Mutagenesis is one of breeding technique for creating gene variability for resistant of disease in soybean. The study aims to examine quantity and quality DNA of soybean mutant lines resistant Athelia rolfsii Curzi. The study was conducted at Plant Biotechnology Laboratory, Faculty of Agriculture, University of Sumatera Utara, Medan. DNA of 7 soybean mutant lines and variety of Anjasmoro was isolated using CTAB extraction method, quantity test using spectrophotometer and quality test using UVITEC Cambridge Fire Reader. The results showed lines that had bright and thick band pattern found in Anjasmoro, M100-25(2/7), M100-A25(3/4), M200-A11(39/7), M300-A8(35/7), while thin and less bright band found in M200-A11(32/3), M200-A12(6/5), M300-A6(33/3). The purity of DNA produced range from 1.84 – 2.00 and the concentration of DNA produced range from 72.9 – 847.3 µg/ml.