Conventional plant breeding usually identifies genetic variation by visual or chemical selection. The process of developing new crop varieties can take up to 25 years. However, with the advancements in molecular biology, the duration has been considerably shortened to 7-10 years. One of the important techniques that makes it efficient for scientists to select plant traits is Marker-Assisted Selection (MAS). In this guide, the most commonly used molecular techniques in tobacco have been surveyed, different types of molecular markers are explained in detail, and recommendations made.