Survival of sperm is limited in the vaginal environment, and DNA of male origin is often partially degraded in vaginal samples obtained from sexual assault cases. This is particularly true for vaginal washes, as vaginal secretions and products of vaginal flora proliferation drained in the wash may contribute to the deterioration of sperm cells after collection of the samples. The time elapsed between the assault and the collection of samples, the forwarding of sexual assault kits to the laboratory and the storage and preparation of specimens for forensic expertise are important factors in the preservation of male DNA. Sexual assault kits received at our laboratory are stored at 4°C until examining samples for the presence of seminal fluid and spermatozoa, after which vaginal washes positive for sperm are stored frozen until DNA extraction. We determined the contribution of both time and storage conditions on the recovery and quality of male DNA in vaginal washes. Fresh sperm was added to three different vaginal washes that were then stored at 4°C. Samples were collected at various time points and processed for differential DNA extraction: 24 hours, 48 hours, 72 hours, 1week and 2 weeks of storage at 4°C. Additional sets of samples were also frozen after each period of storage at 4°C. For each sample, the recovery of male fraction DNA was determined, and the quality of male DNA was evaluated by RFLP analysis on two locus: D4S139 and D10S28. Results will be presented showing the effect of storage at 4°C, freeze-thaw cycle and stringent digestion conditions of the female fraction, on both the loss of male DNA into the female fraction and the degradation of male DNA.