In humans, the brain derived neurotrophic factor (BDNF) gene contains a functional nucleotide single-nucleotide polymorphism (rs6265) which results in a valine to methionine substitution (Val66Met or G 196A), leading to reduced mature BDNF expression. This polymorphism has been widely implicated in a host of disorders and is the focus of many ongoing genetic studies. This study applies the restriction fragment length polymorphism (RFLP) method for identifying BDNF rs6265 polymorphism in Vietnamese. The method is based on the principal that DNA products that result from a polymerase chain reaction are broken into pieces (digested) by restriction enzymes and the resulting restriction fragments are separated according to their lengths by gel electrophoresis. the authors have designed two PCR primers that amplify the BDNF gene region containing rs6265. The digest Pm/l restricted enzyme was selected to detect the G 196A polymorphism. Sequencing the resulting bands validated the RFLP method. In conclusion, this new method describes a sensitive, cost effective and high throughput genotyping of BDNF rs6265 polymorphism, ideal for large-scale genotyping studies.