The reliability of a D1S80 typing procedure has been evaluated using simulated forensic specimens. D1S80 alleles were detectable in DNA recovered from bloodstains exposed to sunlight for up to 20 weeks. However, D1S80 alleles were undetectable in semen stains after six weeks sunlight exposure. Analysis of blood and semen that had been deposited on a variety of substrates and examined over a twenty-week period, revealed no systematic influence of substrate on the ability to type D1S80. A study in which body fluids were exposed to household chemical substances, such as bleach, acids, oil, and gasoline, indicated that only HCl and bleach had a deleterious effect on the ability to type D1S80. In addition, personal care chemical products were without effect on D1S80 allele patterns derived from semen. Exposure of blood and semen to four different species of microorganisms resulted in no alteration of D1S80 genotype patterns in these body fluids. D1S80 genotypes could be reliably determined even when body fluids from different individuals were mixed. DNA from no animals other than humans and higher primates could be amplified at locus D1S80 when the DNA had been isolated through an organic procedure. These studies, in concert with the reports of others, indicate that the procedures for the amplification and detection of genetic variation at locus D1S80 are suitable for use on forensic evidentiary materials.