In this article, we report an efficient method for the synthesis of thyminefunctionalized polystyrene microspheres. First, poly(styrene-co-4-chloromethylstyrene) copolymers slightly crosslinked with divinylbenzene were synthesized in batch free-radical emulsion copolymerization. Microspheres with a particle size of $40-70 nm were obtained with greater than 99% conversion. The chloromethylstyrene (CMS) groups were then converted into thymylmethylstyrene (TMS) in a two-phase system with greater than 80% efficiency, and up to a 45 mol % thymine loading was achieved. The functionalized microspheres were characterized by elemental analysis, Fourier transform infrared, and X-ray photoelectron spectroscopy. The analyses revealed partial hydrolysis of the CMS functionalities, yielding hydroxymethyl functional groups in addition to the thymine functionalities. These copolymers have potential applications in biotechnology. V V C 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 5545-5553, 2005