Anopheles gambiae females in a relatively isolated hut and all larvae from larval habitats within 100 m of the hut were collected in August 2001 in western Kenya. Among 42 aquatic habitats, 16 had A. gambiae larvae. Two hundred fifty larvae and 58 adults were genotyped using nine microsatellite markers to infer sibling relationship between the larvae and maternity between the females and larvae. The pairwise genetic relatedness of A. gambiae larvae per habitat ranged from -0.4112 to 0.9375, indicating that full siblings, half siblings, and genetically unrelated individuals presented at those habitats with multiple larvae. From a likelihood analysis, it was estimated that 56.6% of females had larvae in multiple habitats. These results substantiate that one A. gambiae female uses multiple breeding sites for oviposition, and thus, average genetic relatedness for breeding sites with high larval populations tends to be low.