Related Experiment Video
Updated: Jun 13, 2026
Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
Published on: February 10, 2023
Genotyping the BDNF rs6265 (val66met) polymorphism by one-step amplified refractory mutation system PCR
Haroon I Sheikh1, Elizabeth P Hayden, Katie R Kryski
1Molecular Genetics Unit, Department of Biology, University of Western Ontario, London, Ontario, Canada. hsheikh6@uwo.ca
Background:
The brain derived neutrophic factor (BDNF), a 27 kD polypeptide, is one of the most widely expressed neurotrophins in the brain, regulating neural development and plasticity. The BDNF gene contains a functional single-nucleotide polymorphism (rs6265), which results in a valine to methionine substitution (val66met), leading to reduced mature BDNF expression. This polymorphism has been widely implicated in a host of psychiatric disorders and is a focus of many ongoing psychiatric genetic studies.
Objective:
To develop an efficient and rapid method to detect the val66met polymorphism in a one-step PCR reaction.
Method And Results:
We have designed four PCR primers that amplify the BDNF gene region containing rs6265. The specificity of the four primers in a single PCR reaction amplifies two allele-specific amplicons (253 and 201 bp) and the entire region (401 bp) as an internal control, which are easily distinguished on a polyacrylamide gel. The effectiveness and efficiency of the results are validated by traditional NlaIII restriction enzyme digestion, sequencing of resulting bands and confirmation on 308 genomic DNA samples.
Conclusion:
This new method describes a rapid, sensitive, cost effective and high throughput genotyping of the BDNF val66met polymorphism, ideal for large-scale genotyping studies.
