High levels of homologous recombination are induced during meiosis. This meiotic recombination is initiated by programmed formation of DNA double-strand breaks (DSBs) by a conserved meiosis-specific protein, Spoil. Meiotic DSBs are not formed at random along chromosomes but are formed in clusters known as recombination hot spots. To understand the regulation of this initiation step of meiotic recombination, determining the timing and location of meiotic DSBs is essential. In this chapter, we describe a method to detect genome-wide meiotic DSBs by using a combination of pulsed-field gel electrophoresis and Southern blotting.