Duţă-Cornescu G, Simon-Gruiţă A, Constantin N, Stanciu F, Dobre M, Banică D, Tuduce R, Cristea P, Stoian V. Romanian Biotechnological Letters, 2009;14(6):4845-4850.
Identification of single nucleotide polymorphisms (SNPs) is now possible by many techniques, but choosing one of these methods for a particular case represents quite a challenge, because the researcher must take into consideration many factors.
In this article the authors are trying to present a comparative study of two methods, used currently in our laboratory, for identification of SNPs polymorphisms: ARMS – PCR (amplification refractory mutation system) and RFLP – PCR (restriction fragment length polymorphism). The two SNPs on which we focused belong to human VDR gene (vitamin D receptor gene) and are ApaI (a G→T base change in intron 8) and TaqI (a silent T→C base change in codon 352), named after the restriction enzymes which recognize these variations.
Since the results obtained by both methods were confirmed by direct sequencing, we concluded that ARMS-PCR method is the most adequate for detecting the alternative genotypes determined by single base mutations. The simplicity of this method makes it suitable for the analysis of large number of samples, situation which is usually met in case-control and population genetic studies because this test is easy to use, cost – effective and have an accuracy of 99,9%
Keywords:ARMS-PCR, RFLP-PCR, sequencing, VDR gene, Apa I and Taq I polymorphisms