Fecal samples, tissue paper smeared with fecal material, and clothing that contains human feces, may be the only evidence left at a crime scene. Thus, human stool may be a valuable source for forensic DNA evidence; however, due to microbial presence in the gastrointestinal tracts, the extraction of human DNA from feces may be challenging. Also, stool contains high levels of compounds that can degrade DNA and inhibit downstream PCR reactions; these compounds must be removed for reliable analysis of extracted DNA. In the current study, human stool samples were obtained from eight volunteers, stored under various conditions, and extracted by three different procedures. Fecal material and tissue paper soiled with fecal material obtained from a crime scene were also extracted. Extracted DNA was amplified by using the AmpFlSTR Profiler Plus, AmpFlSTR COfiler, and the AmpFlSTR Identifiler PCR amplification kits for the detection of the autosomal STR allelic patterns. DNA extracted from the male volunteers and from the soiled tissue paper evidence samples was also amplified by using the Y-PLEXtm6 and Y-PLEXtm5 amplification kits. Analysis of the amplified products was conducted by capillary electrophoresis on the ABI PRISM 310 Genetic Analyzer. Autosomal and Y-STR profiles obtained from the fecal material matched the results obtained from the donors' buccal swabs. The study thus found that in cases where fecal matter is the only available evidence, successful profiles may be obtained when the DNA extraction methods described in this report are used. 2 tables, 3 figures, and 11 references