Global climate change has a huge impact on the agricultural world. The water scarcity that happened in some areas can lead to decreased food production, including soybean in Indonesia. Indonesia has a number of soybean genotypes with diverse drought tolerance which have been obtained using various approaches and could be one of the alternatives in responding to the problem. This study aimed to identify soybean mutant genotypes generated from in vitro selections and improved varieties using Simple Sequence Repeat (SSR) markers. A total of 10 mutant lines and 20 improved varieties from Indonesia were genotyped using 10 SSR markers adopted from literatures and ten newly designed SSR markers. The research was conducted in ICABIOGRAD molecular biology laboratory from November 2017 to February 2018. The amplicons were scored as binary data and analyzed using NTSYS and PowerMarker softwares. The new SSR markers were designed based on the variants searched from the IAARD genome database (www.genom.litbang.pertanian.go.id) and showed unambiguous amplicons. The results showed that adopted SSR markers were able to detect more alleles, a higher range of major allele frequency, gene diversity, heterozygosity and Polymorphism Information Content (PIC) compared to the newly designed markers. Phylogenetic analysis showed that all of the soybean mutants were grouped in the same cluster with the parent (Sindoro). This molecular marker-based information of soybean mutants along with the improved varieties in this study could be useful for assisting breeding strategy in screening parental lines to develop drought tolerant soybean varieties in the future.