Insects have long played a role in forensic investigations and can be used to estimate minimum time since death, corpse translocation, and link an individual to a crime scene. Bed bugs (Cimex lectularius) are wingless ectoparasitic insects of potential forensic utility, given that all mobile life stages feed on vertebrate blood. Successful profiling of autosomal short tandem repeats (STRs) from human DNA isolated from bed bugs has been previously reported. This proof-of-concept study looked to expand this work and determine any possible limitations of using bed bugs for both rapid stain identification (RSID™) for human blood and Y-STR profiling. To achieve this, bed bugs were fed either human male only or human pooled (female:male) blood for 30 min and subsequently collected at 12-h intervals up to 108 h post-blood meal (PBM). RSID™ blood testing was successful from the bed bug carcass remaining after DNA isolation, regardless of blood meal type and time of collection PBM. Complete Y-STR profiles were generated from bed bugs <60 h PBM. As the time PBM increased, DNA quantity decreased, while the degradation index increased. Collection of bed bugs at a crime scene could provide a valuable source of human blood for Y STR profiling and be used to link an individual to a crime scene or for potential male suspect exclusion. Future studies should look to replicate the results of this proof-of-concept study with larger numbers of bed bugs, more diverse blood donors, and additional STR profiling kits.