More than 10,000 publications using the random amplified polymorphic DNA (RAPD) or related arbitrary marker techniques have been published in two decades of its inception in 1990. Despite extensive use, RAPD technique has also attracted some criticisms, mainly for lack of reproducibility. In the light of its widespread applications, the objective of this chapter is to (1) provide a protocol for RAPD assay, (2) identify the potential factors affecting the optimization of the RAPD assays, and (3) provide proper statistical analysis to avoid false positives. It is suggested that after proper optimization, the RAPD is a reliable, sensitive, and reproducible assay having the potential to detect a wide range of DNA variations. Analyses of the relevant fragments generated in RAPD profile allow not only to identify some of the molecular events implicated in the genomic instability but also to discover genes playing key roles in genetic evolution and gene mapping. RAPD markers will continue to be boon for genetic studies of those organisms where yet no sequence information or scanty information is available.