Vasectomy is a widely used method of contraception. The absence of spermatozoa in the ejaculate of azoospermic individuals increases the difficulty of extracting DNA for analysis, especially if the biological samples are in the form of spots and stored for years. The aim of this study is to evaluate the possibility of obtaining autosomal and Y-STR profiles from stains containing the ejaculate of individuals pre and postvasectomy, stored in cotton fabric for a period of 10 years. Samples from 28 (twenty eight) individuals: semen pre-vasectomy and seminal fluid (post-vasectomy) stains, stored from 2004 to 2014 at room temperature were used. Three cutouts of 1.5 cm in diameter (punch) were taken from central region of each spot. It was used the QIAamp DNA Mini kit (Qiagen) to extract DNA with an adapted protocol followed by quantification by Nanodrop equipment. Genetic profiles were obtained using ESI17PowerPlexR (Promega). Capillary electrophoresis was performed on the ABI PRISM 3500XL DNA Analyzer. The DNA extraction was successful in all punch analyzed, and its concentration ranged from 1.1 ng/uL to23.3 ng/uL in the pre- vasectomy and from 0.8 ng/uL to 7 ng/uL in postvasectomy punch. The samples had suitable purity, reflecting the presence of few contaminants. Currently, the techniques of DNA amplification are extremely sensitive and capable of assessing human DNA in small quantities generating partial or full profiles, trespassing situations such as long storage periods in cotton fabric. These data can be used as a benchmark for forensic analysis in cases of semen analysis.