Application of restriction fragment length polymorphism method for genotyping tmem18 rs6548238 polymorphism

Le Thi Tuyet, Tran Quang Binh, Duong Thi Anh Dao, Pham Thi Thu Ly


In human, TMEM18 protein (transmembrane protein 18) is involved in the migratory response of neural precursors toward glioma-secreted factors and the overexpression of TMEM18 increased the migration of human neural precursor cells. Single nucleotide polymorphism (SNP) rs6548238 locates at regulatory region near the TMEM18 gene, which results in the substitution of T for C in the DNA sequence. The association of SNP rs6548238 with the risk of several disorders has been intensively studied in developed countries. In order to determine the genotype of TMEM18 rs6548238 polymorphism in Vietnamese, the restriction fragment length polymorphism (RFLP) method was applied. In the RFLP analysis, DNA samples amplified from polymerase chain reaction (PCR) are digested by restriction enzymes and the resulting restriction fragments are separated according to their lengths by gel electrophoresis. In this study, we designed two PCR primers to amplify the region containing rs6548238. The AvaI restriction enzyme was selected to detect the SNP polymorphism. Sequencing the resulting bands validated the RFLP method. In conclusion, RFLP method is simple, sensitive and cost effective for genotyping of the TMEM18 rs6548238 polymorphism; thus, this method is most convenient for SNP analysis in large-size samples from Vietnamese population.


Genotyping, human genetics, restriction enzyme, restriction fragment length polymorphism (RFLP) method, TMEM18-rs6548238

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