A polytene chromosome analysis was prepared from Anopheles freeborni collected from 25 locations in north and central California, and parts of Washington and Oregon. The X chromosome banding pattern, thought previously to be specific to An. hermsi, was common in mosquitoes collected from foothill regions in California, and in all samples from Washington and Oregon. At some of these locations, many mosquitoes had heterokaryotypes for the inversion that distinguishes the X chromosome of An. freeborni from that of An. hermsi. Use of rDNA restriction site analysis, and the results from crossing of different strains bearing either type of X chromosome, showed that An. hermsi does not have a unique or diagnostic X chromosome. Anopheles hermsi was collected in San Mateo County, CA, which is now the northernmost known limit of this species. Crossing studies, or the examination of rDNA restriction enzyme profiles, are presently the only means of identifying An. hermsi.