The revolution in molecular techniques over the last 30 years detracted from many traditional cytological techniques for examining basic biological problems. One of these casualties is the preparation of karyotypes and analysis of chromosomal structure, behaviour, and variation. Recent technology permitting the full sequencing of organisms has highlighted (but does not replace) the importance of understanding chromosomal constitution and karyotype structure, which underpin genome organisation. This chapter provides simple and straightforward protocols for the preparation of chromosome spreads from animals, and more advanced techniques for cell culture and chromosomal banding and hybridisation.